LC3B Antibody

Images

 
Genetic Strategies: Knockout Validated: LC3B Antibody [NB100-2220] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) ...read more
Genetic Strategies: Knockout Validated: LC3B Antibody [NB100-2220] - LC3B/MAP1 [NB100-2220] - Detection of LC3 in mouse ES cell lysates. Atg5-/- ES cells from Dr. Noboru Mizushima [Mizushima, N. et al. J. Cell ...read more
Biological Strategies: Western Blot: LC3B Antibody [NB100-2220] - LC3B/MAP1 [NB100-2220] - Western blot analysis of HeLa (1), HeLa + CQ (2), SHSY5Y (3), SHSY5Y +CQ (4), A431 (5), A431 +CQ (6) and Ntera2 (7) using ...read more
Genetic Strategies: Knockout Validated: LC3B Antibody [NB100-2220] - LC3 was detected in immersion fixed Cloroquine treated Hela cells (left) but was not detected in LC3 knockout Hela cells (right) using rabbit ...read more
Biological Strategies: Western Blot: LC3B Antibody [NB100-2220] - shows lysates of mouse NIH3T3 and rat PC-12 cell lines untreated (-) or treated (+) with Chloroquine. PVDF membrane was probed with 0.5 ug/mL ...read more
Western Blot: LC3B Antibody [NB100-2220] - Analysis of LC3 in Huh-7 and SMMC-7721 cells using anti-LC3B antibody. Image from verified customer review.
Western Blot: LC3B Antibody [NB100-2220] - Analysis using the Biotin conjugate of NB100-2220. Image from verified customer review.
Immunohistochemistry-Paraffin: LC3B Antibody [NB100-2220] - Human ovarian Cancer tissue stained using heat mediated antigen retrieval in pH 6.0 citrate buffer at 1:200 dilution. Image provided by verified customer ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB100-2220] - LC3B detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with Chloroquine using 1ug/mL rabbit anti-LC3B polyclonal ...read more
Western Blot: LC3B Antibody [NB100-2220] - Autophagy Investigation. (A) Representative WBs of LC3B-I, LC3B-II and GAPDH loading control in DANs derived from 4 iPD patients and 4 healthy controls. (B) WBs from three ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB100-2220] - LC3 antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
Immunohistochemistry: LC3B Antibody [NB100-2220] - Analysis using the Biotin conjugate of NB100-2220. Staining of brain, cerebral cortex, neurons with cell processes.
Western Blot: LC3B Antibody [NB100-2220] - Analysis of LC3 in H9C2 cells.
Western Blot: LC3B Antibody [NB100-2220] - Analysis using the DyLight 550 conjugate of NB100-2220. Detection of LC3 in pancreatic cancer cells.
Western Blot: LC3B Antibody [NB100-2220] - Detection of LC3 in mouse HL-1 cell lysate. Photo courtesy of product review by verified customer.
Genetic Strategies: Western Blot: LC3B Antibody [NB100-2220] - WB detection of LC3I and LC3II in mouse cochlea cell line SV-K1. Cells were treated with chloroquine (1uM), and As2O3 (1uM) for 24 hrs. This image ...read more
Western Blot: LC3B Antibody [NB100-2220] - Detection in fibroblast whole cell extract. Image from verified customer review.
Western Blot: LC3B Antibody [NB100-2220] - Macrophage lysates probed with anti-LC3 antibody. This image was submitted via customer Review.
Western Blot: LC3B Antibody [NB100-2220] - Uncropped western blots used for quantification. Detection of LC3B. Normalized Protein levels by the indicated housekeeping protein. iPD genotypes are reported. Blots show ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB100-2220] - HeLa cells were treated with 50uM CQ overnight, fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB100-2220] - Untreated HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells ...read more
Immunohistochemistry: LC3B Antibody [NB100-2220] - IHC of rat brain from confirmed customer review.
Immunohistochemistry-Paraffin: LC3B Antibody [NB100-2220] - IHC analysis of a formalin fixed and paraffin embedded tissue section of mouse brain using 1:200 dilution of rabbit anti-LC3 antibody. The specific signal of ...read more
Immunohistochemistry-Frozen: LC3B Antibody [NB100-2220] - LC3 accumulation in muscle fibers from patients with PAD (row B - E). Normal fibers from a Non-PAD sample (row A). Scale bar = 100 uM. This image was submitted ...read more
Simple Western: LC3B Antibody [NB100-2220] - Simple Western lane view shows a specific band for LC3 in 0.5 mg/ml of Neuro2A lysate. This experiment was performed under reducing conditions using the 12-230 kDa ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Av, Ba, Bv, Ca, Pm, SyHa, ZeSpecies Glossary
Applications WB, Simple Western, ELISA, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, PLA, ChIP, KO
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml

LC3B Antibody Summary

Immunogen
A synthetic peptide made to an N-terminal portion of the human LC3 protein sequence (between residues 1-100). [UniProt# Q9GZQ8]
Localization
LC3-I is cytoplasmic. LC3-II binds to the autophagic membranes.
Marker
Autophagosome Marker
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
MAP1LC3B
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot
  • Simple Western 1:50
  • ELISA
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:200
  • Immunohistochemistry 1:200-1:400
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200-1:400
  • Immunoprecipitation 20ug/500ug of protein
  • Proximity Ligation Assay
  • Chromatin Immunoprecipitation (ChIP)
  • Knockout Validated
Application Notes
This LC3 antibody is tested for ICC/IF, IHC on paraffin embedded sections, IP and Western Blot. Use in IHC on frozen sections reported in scientific literature (PMID: 20008275). Use in immunoblotting reported in scientific literature (PMID 28559895). Use in ELISA reported in scientific literature (PMID 20503249). Use in ChIP reported in scientific literature (PMID28431247). In Western blot bands are seen ~17 and 19 kDa corresponding to LC3-II and LC3-I. In some cases a non-specific band is seen at ~21 kDa in mouse protein. In ICC/IF, cytoplasmic staining was observed in HeLa cells. Use in Proximity Ligation Assay reported in scientific literature (PMID 27219062). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
Control
Brain Cerebral Cortex Lysate (NB820-59182)
Brain Lysate (NB820-59177)
Neuro2a Lysate (NBP2-49688)
HeLa Lysate (NBP2-49689)
Control Peptide
LC3B Peptide (NB100-2220PEP)
Reviewed Applications
Read 39 Reviews rated 4.7
using
NB100-2220 in the following applications:

Publications
Read Publications using
NB100-2220 in the following applications:

  • 1 publication
  • 2 publications
  • 3 publications
  • IB
    4 publications
  • 106 publications
  • IHC
    32 publications
  • 8 publications
  • 14 publications
  • IP
    2 publications
  • PLA
    1 publication
  • WB
    506 publications

Reactivity Notes

Bovine reactivity reported in scientific literature (PMID: 24895572). Primate reactivity reported in scientific literature (PMID: 25142602). Canine reactivity reported in scientific literature (PMID: 25839646). Avian reactivity reported in scientific literature (PMID: 29546310).The mouse detection has been reported to be weaker than the human. Immunogen sequence has 84% homology to Xenopus.

Packaging, Storage & Formulations

Storage
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for LC3B Antibody

  • anti-LC3B
  • Apg8b
  • atg8f antibody
  • ATG8F
  • Autophagy-related protein LC3 B
  • Autophagy-related ubiquitin-like modifier LC3 B
  • lc3b autophagy marker
  • LC3B ihc
  • LC3B immunohistochemistry
  • LC3B immunoprecipitation
  • LC3B western blot
  • LC3B
  • lc3-i, ii
  • LC3II
  • MAP1 light chain 3-like protein 2
  • MAP1A/1BLC3
  • MAP1A/MAP1B LC3 B
  • MAP1LC3B
  • microtubule associated protein 3 b
  • microtubule associated protein 3 beta
  • microtubule-associated protein 1 light chain 3 betaMAP1A/MAP1B light chain 3 B
  • microtubule-associated proteins 1A/1B light chain 3
  • microtubule-associated proteins 1A/1B light chain 3B

Background

LC3 (microtubule-associated protein light chain 3), the most studied autophagy biomarker, was originally identified as a subunit of microtubule-associated proteins 1A and 1B (MAP1LC3) and was later found to contain similarity to yeast protein Apg8/Aut7/Cvt5. Distributed ubiquitously in eukaryotes, LC3 is expressed as 3 splice variants/isoforms (LC3A, LC3B and LC3C) which undergo post-translational processing, wherein, the unprocessed form of LC3 is proteolytically cleaved by Atg4 protease to form LC3-I with carboxyterminal exposed glycine. During autophagy, this exposed glycine of LC3-I is conjugated by Atg7 (an E1-like activity), Atg3 (an E2-like conjugating activity) and by Atg12-Atg5-Atg16L multimers (E3-like ligase activity) to phosphatidylethanolamine (PE) moiety for generating LC3-II. The lipophilic character of PE group facilitates LC3-II insertion into autophagosomes membranes, and as a result LC3-II is degraded when autophagosomes fuse with lysosomes to form autolysosomes for lysus of intra-autophagosomal components by lysosomal hydrolases. Conversion of LC3I to LC3II when correlated with autophagosome numbers is considered as the best marker of autophagy because LC3-II is the only well-characterized protein which specifically localize to autophagic structures throughout autophagy (from phagophore to lysosomal degradation). LC3 is a great tool in research as autophagy is implicated in numerous physiological/pathological processes including responses to exercise/aging, cancer, metabolic and neurodegenerative disorders, and cardiovascular/pulmonary diseases.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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NB100-2220
Species: Hu, Mu, Rt, Po, Av, Ba, Bv, Ca, Pm, SyHa, Ze
Applications: WB, Simple Western, ELISA, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, PLA, ChIP, KO

Publications for LC3B Antibody (NB100-2220)(774)

We have publications tested in 13 confirmed species: Human, Mouse, Rat, Avian, Bacteria, Bovine, Canine, Golden Syrian Hamster, Hamster, Invertebrate, Porcine, Primate, Zebrafish.

We have publications tested in 11 applications: ChIP, ELISA, FLOW, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, PLA, WB.


Filter By Application
ChIP
(1)
ELISA
(2)
FLOW
(3)
IB
(4)
ICC/IF
(106)
IHC
(32)
IHC-Fr
(8)
IHC-P
(14)
IP
(2)
PLA
(1)
WB
(506)
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Human
(312)
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(215)
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(54)
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(1)
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(3)
Bovine
(4)
Canine
(2)
Golden Syrian Hamster
(1)
Hamster
(1)
Invertebrate
(1)
Porcine
(5)
Primate
(2)
Zebrafish
(5)
All Species
Showing Publications 1 - 10 of 774. Show All 774 Publications.
Publications using NB100-2220 Applications Species
Ligeon LA, Loi M, Munz C. LC3-Associated Phagocytosis and Antigen Presentation. Curr Protoc Immunol Sep 25 2018 [PMID: 30253073] (ICC/IF, Human) ICC/IF Human
Brose RD, Lehrmann E, Zhang Y et al. Hydroxyurea attenuates oxidative, metabolic, and excitotoxic stress in rat hippocampal neurons and improves spatial memory in a mouse model of Alzheimer's disease. Neurobiol. Aging Aug 29 2018 [PMID: 30245242] (WB, Rat) WB Rat
Yan J, Porch MW, Court-Vazquez B et al. Activation of autophagy rescues synaptic and cognitive deficits in fragile X mice. Proc. Natl. Acad. Sci. U.S.A. Sep 21 2018 [PMID: 30242133] (WB, Mouse) WB Mouse
Searfoss GH, Paisley BM, Goldstein KM et al. The integrated stress response regulates cell health of cardiac progenitors. Toxicol. Sci. Sep 12 2018 [PMID: 30215789] (WB, Human) WB Human
Auge E, Duran J, Guinovart JJ et al. Exploring the elusive composition of corpora amylacea of human brain. Sci Rep Sep 10 2018 [PMID: 30202002] (IHC, Human) IHC Human
Paul S, Dansithong W, Figueroa KP et al. Staufen1 links RNA stress granules and autophagy in a model of neurodegeneration. Nat Commun Sep 7 2018 [PMID: 30194296] (WB, Human) WB Human
Wong WT, Li LH, Rao YK et al. Repositioning of the B-Blocker Carvedilol as a Novel Autophagy Inducer That Inhibits the NLRP3 Inflammasome. Front Immunol Aug 22 2018 [PMID: 30186288] (WB, Mouse) WB Mouse
Gotzl JK, Colombo AV, Fellerer K et al. Early lysosomal maturation deficits in microglia triggers enhanced lysosomal activity in other brain cells of progranulin knockout mice. Mol Neurodegener Sep 4 2018 [PMID: 30180904] (WB, Mouse) WB Mouse
Lin CW, Chen YS, Lin CC et al. Significant predictors of overall survival in patients with hepatocellular carcinoma after surgical resection. PLoS ONE Sep 4 2018 [PMID: 30180193] (IHC, Human) IHC Human
Kaur G, Gauthier SA, Perez-Gonzalez R et al. Cystatin C prevents neuronal loss and behavioral deficits via the endosomal pathway in a mouse model of down syndrome. Neurobiol. Dis. Sep 1 2018 [PMID: 30176349] (WB, Mouse) WB Mouse
Show All 774 Publications.

Reviews for LC3B Antibody (NB100-2220) (39) 4.739

Average Rating: 4.7
(Based on 39 reviews)
We have 39 reviews tested in 6 species: Human, Mouse, Rat, Mouse and Human, Other, murine.

Reviews using NB100-2220:
Filter by Applications
WB
(31)
ICC
(1)
IHC-Fr
(1)
IF
(1)
Simple Western
(2)
IHC
(1)
IHC-P
(1)
IP
(1)
All Applications
Filter by Species
Human
(16)
Mouse
(13)
Rat
(4)
Mouse and Human
(2)
Other
(2)
murine
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Human 07/17/2018
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Summary

ApplicationWestern Blot
Sample TestedTHP-1 whole cell lysate
SpeciesHuman
LotAQ-1

Comments

CommentsTHP-1 cells were treated with Rapamycin (100nm) for 24h. Total cell lysates were subjected to western blot. PVDF membrane were probed with 1mm/ml Human LC3B Antibody (NB100-2220) Antibody. A specific band was detected for LC3B I and LC3B II at approximately 13~15 kDa. This experiment was conducted under reducing conditions
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Human 06/07/2018
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Summary

ApplicationWestern Blot
Sample Testedwhole cell lysate,fibroblasts whole cell extract
SpeciesHuman
LotDJ-1
Immunocytochemistry LC3B NB100-2220
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reviewed by:
Anonymous
ICC Human 05/18/2018
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Summary

ApplicationImmunocytochemistry
Sample Tested hela cell,HEK 293
SpeciesHuman
LotDK-2
Western Blot LC3B NB100-2220
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reviewed by:
Dongkai Guo
WB Mouse and Human 05/10/2018
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Summary

ApplicationWestern Blot
Sample TestedNIH3T3, HeLa, HEK293T, N2a neuroblastoma, Q7 striatal, Sy5Y neuroblastoma
SpeciesMouse and Human
LotDK-2
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Mouse 02/22/2018
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Summary

ApplicationWestern Blot
Sample Testedcochlea
SpeciesMouse
LotDQ-1
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Mouse and Human 11/25/2017
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Summary

ApplicationWestern Blot
Sample Testedmouse hepatocytes and HepG2 cells
SpeciesMouse and Human
LotDM-1
Immunohistochemistry-Frozen LC3B NB100-2220
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reviewed by:
Kate Kosmac
IHC-Fr Human 08/23/2017
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Summary

ApplicationImmunohistochemistry-Frozen
Sample TestedHuman skeletal muscle
SpeciesHuman
LotCX

Comments

CommentsFor LC3: Fresh-frozen sections were acetone fixed, washed with TBST(1% Tween), blocked for 1 hr with TBST, 5% NGS and incubated ON at 4C with primary antibody. LC3 staining was amplified with biotin followed by a directly conjugated streptavidin.
 
reviewed by:
Anonymous
WB Rat 07/14/2017
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Summary

ApplicationWestern Blot
Sample TestedNRK rat normal kidney cell line
SpeciesRat

Comments

CommentsUsed at 1:2500. Please see publication (Lee WK et al. Arch Toxicol 2017, in press) for details.

NOVUSCITED
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Human 04/12/2017
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Summary

ApplicationWestern Blot
Sample Tested20 ug whole cell lysate
SpeciesHuman
LotDH-2
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Mouse 03/23/2017
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Summary

ApplicationWestern Blot
Sample TestedMacrophage
SpeciesMouse
LotDG-1
 
reviewed by:
Paola Menichini
WB Human 03/14/2017
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Summary

ApplicationWestern Blot
Sample TestedMDA MB 231 whole cell lysates and conditioned medium
SpeciesHuman
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB murine 02/14/2017
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Summary

ApplicationWestern Blot
Sample Testedmurine bone marrow derived macrophages
Speciesmurine
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Rat 01/18/2017
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Summary

ApplicationWestern Blot
Sample TestedChondrocyte cell lysate
SpeciesRat
LotCL
Immunofluorescence LC3B NB100-2220
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reviewed by:
Anonymous
IF Mouse 01/29/2016
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Summary

ApplicationImmunofluorescence
Sample Testedmouse HSPCs
SpeciesMouse

Blocking

Blocking Details5% fetal bovine serum in PBS for 1 h at RM.

Primary Anitbody

Dilution Ratio1:1000, overnight, 4-C, in PBS.

Secondary Antibody

Secondary Descriptionrhodamine (red) conjugated donkey anti-rabbit IgG (sc-2095)
Secondary Manufacturer Cat#sc-2095
Secondary Concentration0.4ug/ml

Details

Detection NotesOlympus IX71
Fixation DetailsCells were fixed by 4% PFA in PBS for 10 min at RT and then permeabilized in 0.5% Triton X-100 PBS for 10 min.
Wash Description0.5% PBST 5min X 3 times
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Mouse 01/29/2016
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Summary

ApplicationWestern Blot
Sample Testedmouse HSPCs
SpeciesMouse

Blocking

Blocking Details5% fat-free milk in PBS

Primary Anitbody

Dilution Ratio1:5000, overnight, 4C, 5% fat-free milk in PBST (0.5%TWEEN-20)

Secondary Antibody

Secondary Descriptiongoat anti-rabbit IgG-HRP
Secondary Manufacturer Cat#sc-2004
Secondary Concentration0.2ug/ml

Details

Detection NotesECL, EXPOSEURE TIME:30s
Simple Western LC3B NB100-2220
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reviewed by:
Anonymous
Simple Western Mouse 01/21/2016
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Summary

ApplicationSimple Western
Sample Testedmouse brain tisseu lysate
SpeciesMouse

Blocking

Blocking Detailsn/a

Primary Anitbody

Dilution Ratio1:200

Details

Detection NotesWes Simple Western platform
Simple Western LC3B NB100-2220
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reviewed by:
Anonymous
Simple Western Human 01/21/2016
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Summary

ApplicationSimple Western
Sample TestedSH-SY5Y cell lysate
SpeciesHuman

Blocking

Blocking Detailsn/a

Primary Anitbody

Dilution Ratio1:200

Details

Detection NotesWes Simple Western platform
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Mouse 01/20/2016
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Summary

ApplicationWestern Blot
Sample TestedMouse eye tissue
SpeciesMouse

Blocking

Blocking DetailsBlocking with 5% milk in TBST at room temperature for 1 hr

Details

Detection NotesLC3 antibody was diluted by 1000 times. ECL was used as substrate. Detect with ImageQuant.
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Human 07/25/2015
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Summary

ApplicationWestern Blot
Sample TestedHuman prostate cancer cell line (PC3)
SpeciesHuman

Blocking

Blocking Details5% milk in TBS buffer supplemented with 0.1% tween20 at room temperature for one hour

Primary Anitbody

Dilution Ratio1:10000

Secondary Antibody

Secondary DescriptionAnti-Rabbit secondary diluted at 1:10000 in 5% milk
Secondary Manufacturer Cat#SC-2004
Secondary Concentration0.4 mg/ml

Details

Detection NotesWestern blotting using ECL reagent (purchased from GE life sciences) and x-ray films.
Western Blot LC3B NB100-2220
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reviewed by:
Chunhui Sun
WB Human 01/30/2015
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Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate
SpeciesHuman

Blocking

Blocking DetailsBSA, 5%, 1 hour, RT

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG

Details

Detection NotesECL
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Other 01/20/2015
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Summary

ApplicationWestern Blot
Sample TestedGuinea pig ling, liver and heart
SpeciesOther

Blocking

Blocking DetailsMembrane Blocking Solution, Cat# 00-0105 (Life Technologies)

Primary Anitbody

Dilution Ratio1:500

Secondary Antibody

Secondary DescriptionAnti-rabbit IgG, HRP-linked Antibody #7074 (Cell Signaling Technology)
Secondary Manufacturer Cat##7074
Secondary Concentration1:1000

Details

Detection Notes5 seconds
Western Blot LC3B NB100-2220
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reviewed by:
Liyong Zhang
WB Mouse 06/26/2014
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Summary

ApplicationWestern Blot
SpeciesMouse
FileView PDF
 
reviewed by:
Lindsy Dickerson
WB Mouse 06/13/2014
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Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
Western Blot LC3B NB100-2220
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reviewed by:
Anonymous
WB Human 03/27/2014
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Summary

ApplicationWestern Blot
SpeciesHuman
LotBK-1
FileView PDF
Immunohistochemistry LC3B NB100-2220
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reviewed by:
Ashwini Katre
IHC Human 02/28/2014
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Summary

ApplicationImmunohistochemistry
Sample TestedOvarian cancer tissue
SpeciesHuman
LotBJ-1

Blocking

Blocking Details10% goat serum room temp 1 hour

Primary Anitbody

Dilution Ratio1:200 in 10% goat serum

Secondary Antibody

Secondary Descriptiongoat anti rabbit HRP polymer from vector labs

Details

Detection NotesDAB for 3 min
Fixation DetailsAntigen retrieval in pH6 citrate buffer pressure cook for 5 min
Wash Description1XPBS 3minX3
Immunohistochemistry-Paraffin LC3B NB100-2220
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reviewed by:
Anonymous
IHC-P Rat 01/16/2014
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Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedRat brain
SpeciesRat
Western Blot LC3B NB100-2220
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reviewed by:
Angela NIE
WB Mouse 02/12/2013
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Summary

ApplicationWestern Blot
Sample Testedmouse ventricle
SpeciesMouse

Blocking

Blocking Details5%milk TBST

Primary Anitbody

Dilution Ratio5%BSA TBST

Secondary Antibody

Secondary Descriptionanti-rabbit

Details

Detection NotesECL
Western Blot LC3B NB100-2220
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reviewed by:
Myriam Elschami
WB Mouse 12/12/2012
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Summary

ApplicationWestern Blot
Sample TestedHL-1 cell lysate
SpeciesMouse
CommentsThe antibody very specifically stains LC3-I and LC3-II, nicely allowing for the quantification of autophagy by western blotting.

Blocking

Blocking DetailsOdyssey Blocking buffer/PBST 1:1; 1h Room temperature

Primary Anitbody

Dilution Ratio1:1000 in Odyssey Blocking buffer/PBST 1:1, over night, 4°C

Secondary Antibody

Secondary Descriptiongoat anti rabbit IRDye 800CW
Secondary Manufacturer Cat#Li-Cor # 926-32211
Secondary Concentration1:20000 of 1mg/ml stock

Details

Detection NotesLicor Imaging

Comments

CommentsThe antibody very specifically stains LC3-I and LC3-II, nicely allowing for the quantification of autophagy by western blotting.
 
reviewed by:
Tania Rogan
WB 08/10/2012
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Summary

ApplicationWestern Blot
Special ApplicationsI have tried several LC3 antibodies including ones from MBL and Cell Signaling. Novus' antibody was by far most sensitive, and probably cleaner in terms of number and intensity of the non-specific bands it reacted with. Out of 3 antibodies I ended up using Novus' antibody for staining in vitro and in vivo samples.
FileView PDF
Western Blot LC3B NB100-2220
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reviewed by:
Angela NIE
WB Rat 01/09/2012
View

Summary

ApplicationWestern Blot
Sample TestedH9C2 extract, Sample Amount: 20ug
SpeciesRat
LotA-K
CommentsThe testing results were good. I am pleased with this Antibody.

Blocking

Blocking DetailsBlocking Buffer: 5% Milk in TBST, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

Dilution RatioDilution Ratio: 1:1000, Incubation Dilution Buffer: TBST, Incubation Time: 4C,O/N, Incubation Temp: 4 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab: Donkey anti Rabbit (GE Healthcare Life Sciences #NA934V)
Secondary Manufacturer Cat#na934v,GE health

Details

Detection NotesDetection Method: ECL Plus, Exposure Time: 3 seconds, Specific Bands: 17kd kDa

Comments

CommentsThe testing results were good. I am pleased with this Antibody.
 
reviewed by:
Anonymous
WB Human 12/05/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
CommentsClear

Blocking

Blocking DetailsBlocking Buffer: FBS

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:5000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab Dilution Ratio: 1:5000

Details

Detection NotesDetection Method: ECL

Comments

CommentsClear
 
reviewed by:
Anonymous
WB Mouse 11/21/2011
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
CommentsClear

Blocking

Blocking DetailsBlocking Buffer: 5% milk, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:500, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab Dilution Ratio: 1:20000

Details

Detection NotesDetection Method: HRP ECL

Comments

CommentsClear
 
reviewed by:
Anonymous
WB Human 11/21/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman

Blocking

Blocking DetailsBlocking Buffer: 5% non-fat milk in TBST

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:500, Primary Ab Incubation Time: overnight

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-rabbit

Details

Detection NotesDetection Method: Western Lighting plus ECL
 
reviewed by:
Anonymous
WB Mouse 11/16/2011
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse

Blocking

Blocking DetailsBlocking Buffer: BSA

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-rabbit HRP

Details

Detection NotesDetection Method: ECL
 
reviewed by:
Anonymous
WB Human 11/03/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman

Blocking

Blocking DetailsBlocking Buffer: 5% Milk

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:8000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab Dilution Ratio: 1:10000

Details

Detection NotesDetection Method: ECL
 
reviewed by:
Anonymous
WB Human 10/31/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
CommentsClear and significant!

Blocking

Blocking DetailsBlocking Buffer: 5% Milk in TBST

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: overnight, Primary Ab Incubation Temp: 4 degrees Celsius

Details

Detection NotesDetection Method: ECL

Comments

CommentsClear and significant!
 
reviewed by:
Anonymous
WB Human 10/31/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman

Blocking

Blocking DetailsBlocking Buffer: 5% non-fat milk

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:2000, Primary Ab Incubation Time: overnight

Secondary Antibody

Secondary DescriptionSecondary Ab: Goat-anti Rabbit

Details

Detection NotesDetection Method: ECL
 
reviewed by:
Devin Wolfe
WB Mouse 07/21/2010
View

Summary

ApplicationWestern Blot
Sample TestedMurine Blastocyst Derived Cells, Sample Amount: 30ug
SpeciesMouse
CommentsThis particular antibody worked beautifully for WB, with one minor caveat. After repeated usage, the binding of the antibody to LC3-I seems to drop off significantly vs. LC3-II, making quantification of LC3 levels in reference to each isoform significantly more challenging, and borderline impossible due to these fluctuations. Again, was specific, looked good, but lost potency for LC3-I after repeated usage.

Blocking

Blocking DetailsBlocking Buffer: 5% milk in TBST, Blocking Time: 1 hour, Blocking Temp: 25 degrees Celsius

Primary Anitbody

Dilution RatioDilution Ratio: 1:1000, Incubation Dilution Buffer: 1% BSA in TBST, Incubation Time: 1 hour, Incubation Temp: 25 C

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-rabbit HRP conjugated, Secondary Ab Dilution Ratio: 1:5000

Details

Detection NotesDetection Method: Hydrogen Peroxide ECL kit, Exposure Time: 5 minutes, Specific Bands: 15 and 17 kDa

Comments

CommentsThis particular antibody worked beautifully for WB, with one minor caveat. After repeated usage, the binding of the antibody to LC3-I seems to drop off significantly vs. LC3-II, making quantification of LC3 levels in reference to each isoform significantly more challenging, and borderline impossible due to these fluctuations. Again, was specific, looked good, but lost potency for LC3-I after repeated usage.
 
reviewed by:
Anonymous
IP Other 06/15/2009
View

Summary

ApplicationImmunoprecipitation
Sample TestedIP Sample
SpeciesOther
CommentsI used both NB100-2220ss and NB100-2331ss for IP, NB100-2220ss works very well, but not NB100-2331ss. Detected with NB600-1384. Lysate Amount: 0.25mg, Ab Dilution Ratio: 20ug/250ug lysate, Protein Capture: Protein A + Protein B, Protein Reduction: RIPA buffer

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: 3 hours, Primary Ab Incubation Temp: 4 degrees Celsius

Comments

CommentsI used both NB100-2220ss and NB100-2331ss for IP, NB100-2220ss works very well, but not NB100-2331ss. Detected with NB600-1384. Lysate Amount: 0.25mg, Ab Dilution Ratio: 20ug/250ug lysate, Protein Capture: Protein A + Protein B, Protein Reduction: RIPA buffer

Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for LC3B Antibody (NB100-2220). (Showing 1 - 8 of 8 FAQs).

  1. What are the best blocking conditions for NB100-2220?
    • The conditions we have had the most success with are 5% milk + 1% BSA as the block. Our lab also recommends doing an overnight incubation as this is how we run our QC tests in the lab at 4 degrees C.
  2. Is NB100-2220 the same or different from NBP-48614?
    • NB100-2220 is an anti-LC3 Ab. It is similar to NBP1-48614, however they are different Abs. NB100-2220 is specific for only LC3B where as NBP1-48614 is specific for only LC3A I would recommend NB100-2220 if you would like to detect LC3B as it has been in 48 published papers and it has very good reviews. NB100-2220 does detect both LCI and LCII.
  3. What proteins are recognized by this antibody? On the datasheet, the Specificity section states 'This antibody is specific for LC3B.' However, in the Gene Symbol is listed as MAP1LC3A and the protein sequence Q9H492 is for MAP1LC3A. Can you tell me which target the antibody recognizes?
    • This antibody was designed to detect both LC3I and LC3II, but we have experienced through our testing that it much more routinely picks up LC3B. It will sometimes pick up both bands, but it depends on how the cells are prepared often times if they are induced to express that second LC3A band, which is why we have it named as LC3B. We have other LC3 antibodies that do pick up both bands regularly.
  4. What is the isotype of this antibody? I would like to find an isotype control.
    • This is a polyclonal antibody from rabbit. The isotype of this antibody, and all rabbit antibodies, will be IgG. A list of our rabbit secondary antibodies can be found at this link.
  5. Does this antibody also recognizes LC3A and LC3C?
    • This antibody was raised using a synthetic peptide made to an N-terminal portion of the human LC3 protein sequence as immunogen and is expected to detect LC3B protein. The immunogenic sequence employed does not have any similarity to LC3A or LC3C and hence we do not expect any cross-reactivity. This product is one of our best sellers with great customer reviews, as well as with citations in at least 160 peer reviewed publications.
  6. Hello,I already have the antibody, but I was wondering if you could tell me the percent identity to a specific zebrafish LC3 protein. I know it says it works in zebrafish, but we have three and the one I am interested in is the one expressed maternally--- F1QKC7. Here is the Alignment to the human protein your antibody was made. Would you just look at it quick and tell me if the 10 aa peptide is one that is highly similar (like for example aa's 66-71). http://www.uniprot.org/align/2013102541KABHU792. LC3 Antibody (NB100-2220) It is very closely related to my maternal expressed protein. I was just wanting to confirm that it is the right thing and not background staining that I am seeing.
    • Based on the actual immunogen sequence, I am only seeing a 31% homology to the zebrafish sequence you have provided. While the homology is low, there is still a possibility that the antibody could recognize the zebrafish protein. Just for your information, the zebrafish sequence we used for validation was accession number Q7ZUD8. I can offer a few suggestions to verify specific staining in your samples: If it is possible, I would verify by western blot with your tissue. If you can validate by two applications, the staining seen is most likely specific. With the staining, I would run a secondary only or isotype control to ensure that any background staining is not coming from the secondary antibody or other interaction. If you do have access to a knock-down or knock-out, this would be the most ideal way to verify specific staining.
  7. We just bought the LC3 antibody (NB100-2220) from your company, and this product arrived at our lab today. Unfortunately, I just found that the label on the antibody vial showed "pAB LC3 antibody", so we are confused with the "p", what does "P"mean, usually it means"phosphorylated". So, I am wondering what does "p" mean, as the phosphorylated antibody is not we wanted. BTW, the datasheet of this product never mentioned the "phosphorylated"
    • I am sorry for any confusion. "pAB" stands for polyclonal antibody. Our NB100-2220 is not phosphorylated.
  8. I want to know how the lc3 antibody (nb100-2220) was produced and purified?
    • Of course. It is immunogen affinity purified. The immunogen is A synthetic peptide made to an N-terminal portion of the human LC3 protein sequence (between residues 1-100)

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB100-2220AF405
Alexa Fluor 488 Labeled NB100-2220AF488
Alexa Fluor 647 Labeled NB100-2220AF647
Alexa Fluor 700 Labeled NB100-2220AF700
Biotin Labeled NB100-2220B
DyLight 405 Labeled NB100-2220V
DyLight 488 Labeled NB100-2220G
DyLight 550 Labeled NB100-2220R
DyLight 650 Labeled NB100-2220C
FITC Labeled NB100-2220F
HRP Labeled NB100-2220H

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Blogs on LC3B. Showing 1-10 of 12 blog posts - Show all blog posts.

Epigenetic Control of Autophagy
By Christina Towers, PhD. In the last 20 years, epigenetic regulation has become front and center for almost all fields of biology and its role in diseases like cancer and neurodegeneration are being heavily studied....  Read full blog post.

Crosstalk Between Oxidative Stress and Autophagy
By Christina Towers, PhD. Role of Reactive Species in Cellular FunctionOxidative stress is a byproduct of an imbalance between oxidants and antioxidants present in the cell resulting in dysfunctional redox signali...  Read full blog post.

Autophagy inhibition in pediatrics: One physician-scientist’s brave decision
By Christina Towers, PhD. The current time from when a discovery is first made on the bench to when that discovery might translate into an approved therapy in cancer patients is an astounding 10-15 years. Scientists...  Read full blog post.

Cleaner gone bad: Autophagy regulates motor neuron loss in spinal muscular atrophy
By Jamshed Arslan Pharm.D. Neuromuscular disorders affect the peripheral nervous system and muscles. Spinal muscular atrophy (SMA) is one such incurable disease in which muscles fail to receive signals from the spi...  Read full blog post.

Brain size matters: MTOR regulates autophagy and number of cortical interneurons
By Jamshed Arslan Pharm.D. Interneurons transmit impulses between other neurons, in part, to facilitate the birth of neurons. Cortical interneurons themselves arise from the progenitors in the ventral telencephalon,...  Read full blog post.

Why LC3B Antibodies Make Ideal Autophagosomes Membrane Markers
The human form of microtubule-associated protein light chain 3 (LC3) is expressed as 3 splice variants; LC3A, LC3B and LC3C (He et al., 2003). LC3B is a subunit of the MAP1A and MAP1B microtubule-binding pr...  Read full blog post.

LC3B: From Autophagy to Cancer
LC3B is subunit component of the LC3 autophagy biomarker associated with microtubule-associated proteins MAP1A and MAP1B and one of the best characterized markers to date. In resting state it is cytosolic, but upon activation, is lapidated and becomes...  Read full blog post.

LC3B Empowers Protein Quality Control by Autophagy
LC3B also known as microtubule-associated protein 1 light chain 3 beta (MAP1LC3B) is an autophagy gene that contributes appreciably to protein degradation. Autophagy is a highly synchronized and a dynamic catabolic degradation activity that plays an e...  Read full blog post.

Analyzing LC3 in Western blot
Microtubule-associated protein light chain 3 (LC3) is considered one of the definitive markers of autophagy, and its use is widespread in labs throughout the world. Despite its popularity, there are several considerations when employing LC3 antibodies...  Read full blog post.

Marking the Autophagosome: the LC3 Antibody
MAP1LC3 (shortened to LC3 in our antibody catalog) is one of four mammalian homologues of autophagy-related protein 8 (Atg8). It has been identified as a light chain subunit of the microtubule-associated proteins MAP1A/MAP1B. A modified form of LC3, L...  Read full blog post.

Showing 1-10 of 12 blog posts - Show all blog posts.

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Recent Reviews

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Anonymous
07/17/2018
Application: WB
Species: Human

Anonymous
06/07/2018
Application: WB
Species: Human

Anonymous
05/18/2018
Application: ICC
Species: Human

Bioinformatics

Gene Symbol MAP1LC3B