LC3B Antibody

Images

 
Genetic Strategies: Knockout Validated: LC3B Antibody [NB100-2220] - Lysates of HeLa parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 uM Chloroquine for 18 hours. ...read more
Genetic Strategies: Knockout Validated: LC3B Antibody [NB100-2220] - LC3B was detected in immersion fixed Cloroquine treated HeLa cells (left) but was not detected in LC3B knockout HeLa cells (right) using rabbit ...read more
Western Blot: LC3B Antibody [NB100-2220] - Autophagy Investigation. (A) Representative WBs (Western Blots) of LC3B-I, LC3B-II and GAPDH loading control in DANs derived from 4 iPD patients and 4 healthy controls. (B) WBs ...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. Image from verified ...read more
Immunohistochemistry: LC3B Antibody [NB100-2220] - Localization of LC3 by immunohistochemical method. The immunohistochemical staining shows that LC3, CRF, and HIF-1alpha have an overlapping localization in villous and ...read more
Western Blot: LC3B Antibody [NB100-2220] - Detection of LC3B in mouse ES cell lysates using Rabbit anti- [Catalog # NB100-2220]. LC3 detected at a molecular weight of approximately 17 kDa. Atg5-/- ES cells from Dr. ...read more
Western Blot: LC3B Antibody [NB100-2220] - Autophagy induced by ceramide in CNE2 and SUNE1 cells. Ceramide dose and time-dependently induced the formation of LC3-II, a marker for autophagy. SUNE1 cells were treated with ...read more
Western Blot: LC3B Antibody [NB100-2220] - GNA triggers the formation of autophagic markers in A549 and HeLa cell. Effects of GNA on LC3 protein. A549 cells were treated with various concentrations of GNA for 24 hours ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB100-2220] - ORP1L and VAP-A form cholesterol-dependent ER-AV contact sites MelJuSo cells expressing GFP or GFP-tagged ORP1L mutants were fixed and stained for ...read more
Immunohistochemistry: LC3B Antibody [NB100-2220] - SCI leads to lysosomal dysfunction. Representative high magnification images ( x 60) of LC3 (green) and CTSD (red) staining in VH of gray matter from sham and day 1 SCI ...read more
Biological Strategies: Western Blot: LC3B Antibody [NB100-2220] - Lysates of mouse NIH3T3 and rat PC-12 cell lines untreated (-) or treated (+) with Chloroquine. PVDF (Polyvinylidene difluoride) membrane was ...read more
Biological Strategies: Western Blot: LC3B Antibody [NB100-2220] - Analysis of LC3 in untreated and treated H9C2 cells using Rabbit anti-LC3B Antibody [Catalog # NB100-2220].
Biological Strategies: Western Blot: LC3B Antibody [NB100-2220] - Detection of LC3I and LC3II in mouse cochlea cell line SV-K1 using Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Cells were treated with ...read more
Biological Strategies: Western Blot: LC3B Antibody [NB100-2220] - Expression of autophagic protein LC3 in BME-UV1 cells forming acinar structures on Matrigel. Representative images of Western blot analysis of LC3 ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB100-2220] - Autophagosomes accumulate in neurons of the VH gray matter at day 1 after SCI. Representative images of IHC staining for LC3 (green) and neuronal ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB100-2220] - Autophagosomes accumulate in microglia and oligodendrocytes of the dorsal white matter adjacent to injury after SCI. Representative images of IHC ...read more
Immunohistochemistry: LC3B Antibody [NB100-2220] - Staining of rat brain tissue section. Analysis using Rabbit anti-LC3B Antibody [Catalog # 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
Immunohistochemistry-Paraffin: LC3B Antibody [NB100-2220] - FFPE (Formalin-Fixed Paraffin-Embedded) tissue section of mouse brain using 1:200 dilution of Rabbit anti-LC3B antibody [Catalog # NB100-2220]. The specific ...read more
Immunohistochemistry: LC3B Antibody [NB100-2220] - Analysis using the Biotin conjugate of Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Staining of brain, cerebral cortex, neurons with cell processes.
Simple Western: LC3B Antibody [NB100-2220] - Image shows a specific band for LC3B at a molecular weight of approximately 15 kDa in 0.5 mg/mL of Neuro2A lysate. This experiment was performed under reducing conditions ...read more

Product Details

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

LC3B Antibody Summary

Immunogen
Polyclonal LC3B Antibody was made to a synthetic peptide made to an N-terminal portion of the human LC3B protein sequence (between residues 1-100). [UniProt# Q9GZQ8]
Localization
Type I form of LC3B is cytoplasmic, whereas the type II form of LC3B 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 0.5 - 2.0 ug/mL
  • Simple Western 1:50
  • ELISA
  • Flow Cytometry
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:200
  • Immunohistochemistry 1:200 - 1:400
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200 - 1:400
  • Immunoprecipitation 20 ug/500 ug of protein
  • Proximity Ligation Assay
  • Chromatin Immunoprecipitation (ChIP)
  • Knockdown Validated
  • Knockout Validated
Application Notes
In Western blot, bands are seen at ~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 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 (PMID: 28431247). Use in Proximity Ligation Assay reported in scientific literature (PMID: 27219062). Use in FLOW reported in scientific literature (PMID: 27980456). Use in IHC on paraffin-embedded sections reported in scientific literature (PMID: 31071489). Use in Immunoprecipitation reported in scientific literature (PMID: 31209065). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Knockdown validation (PMID: 31754102).
Theoretical MW
14.688 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Control
Human Brain Cerebral Cortex Whole Tissue Lysate (Adult Whole Normal)
Human Brain Whole Tissue Lysate (Adult Whole Normal)
Neuro2a Chloroquine Treated / Untreated Cell Lysate
HeLa Chloroquine Treated / Untreated Cell Lysate
Control Peptide
LC3B Protein (NB100-2220PEP)
Reviewed Applications
Read 47 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
  • EM
    1 publication
  • 3 publications
  • IB
    8 publications
  • 162 publications
  • IHC
    44 publications
  • 11 publications
  • 25 publications
  • IP
    7 publications
  • KD
    4 publications
  • KO
    4 publications
  • PLA
    1 publication
  • WB
    721 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). Hamster reactivity reported in scientific literature (PMID: 26423766). Rabbit reactivity reported in scientific literature (PMID: 26497211). The mouse detection has been reported to be weaker than the human. Immunogen displays the following percentage of sequence identity for non-tested species: Xenopus 84%. Invertebrate reactivity reported in scientific literature (PMID: 26716072 ). Monkey reactivity reported in scientific literature (PMID: 30324853). Guinea pig reactivity reported from a verified customer review. Rat reactivity reported in scientific literature (PMID: 30744518). Bacteria reactivity reported in scientific literature (PMID: 28783414). Chicken reactivity reported in scientific literature (PMID: 30649814). Porcine reactivity reported in scientific literature (PMID: 30789643). Golden syrian hamster reactivity reported in scientific literature (PMID: 23180219). Zebrafish reactivity reported in scientific literature (PMID: 29185873).

Packaging, Storage & Formulations

Storage
Store at -20C.
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
  • MAP1LC3B-a
  • microtubule associated protein 1 light chain 3 beta
  • microtubule associated protein 3 b
  • microtubule associated protein 3 beta
  • microtubule-associated proteins 1A/1B light chain 3
  • microtubule-associated proteins 1A/1B light chain 3B

Background

Autophagy (macroautophagy) is a catabolic process which targets intracellular components such as proteins and organelles for degradation. Originally described as a bulk degradation process, current research supports its selective nature (1). Selective autophagy targets specific cellular components for degradation including the endoplasmic reticulum (2) (ER-phagy), mitochondria (3) (mitophagy), peroxisomes (3) (pexophagy), ribosomes (4) (ribophagy) and bacteria (5) (xenophagy). Autophagy relies on a newly formed phagophore, a membrane structure which elongates, sequesters cellular content, and fuses to form a double membrane vesicle known as the autophagosome. Fusion of autophagosomes with lysosomes gives rise to the autophagolysosome, where cellular components are degraded by lysosome hydrolases (1).

Autophagic flux is supported by autophagy-related proteins (Atgs) initially identified in yeast (6,7). The core autophagy machinery is comprised of 17 Atg proteins that play specific roles in autophagosome formation. Among these Atg proteins, Atg8 is not only involved in autophagosome formation but also functions in cargo selection. In mammals, several Atg8 homologues have been identified including microtubule-associated protein 1 light chain 3 alpha, beta and gamma - LC3A, LC3B, and LC3C (8) respectively, as well as GABA type A receptor-associated protein (GABARAP), GABARAP-Like1, and GABARAP-Like2 (9). LC3 (predicted molecular weight 14kD) is ubiquitously expressed and undergoes posttranslational processing after synthesis. First, the cysteine protease Atg4 cleaves a carboxy terminal sequence to generate the cytosolic form LC3-I. Next, E1-like (Atg7) and E2-like (Atg3) enzymes conjugate phosphatidylethanolamine to the newly exposed carboxyterminal glycine, generating LC3-II. Finally, the Atg12-Atg5-Atg16L1 complex participates in LC3 lipidation and autophagosome formation (10). LC3B-I to LC3B-II conversion correlates with autophagosome number and is considered the best marker to monitor autophagy.

References

1. Yu, L., Chen, Y., & Tooze, S. A. (2018). Autophagy pathway: Cellular and molecular mechanisms. Autophagy. https://doi.org/10.1080/15548627.2017.1378838

2. Forrester, A., De Leonibus, C., Grumati, P., Fasana, E., Piemontese, M., Staiano, L.,... Settembre, C. (2019). A selective ER -phagy exerts procollagen quality control via a Calnexin- FAM 134B complex. The EMBO Journal. https://doi.org/10.15252/embj.201899847

3. He, X., Zhu, Y., Zhang, Y., Geng, Y., Gong, J., Geng, J.,... Zhong, H. (2019). RNF34 functions in immunity and selective mitophagy by targeting MAVS for autophagic degradation. The EMBO Journal. https://doi.org/10.15252/embj.2018100978

4. Mathai, B., Meijer, A., & Simonsen, A. (2017). Studying Autophagy in Zebrafish. Cells. https://doi.org/10.3390/cells6030021

5. Losier, T. T., Akuma, M., McKee-Muir, O. C., LeBlond, N. D., Suk, Y., Alsaadi, R. M.,... Russell, R. C. (2019). AMPK Promotes Xenophagy through Priming of Autophagic Kinases upon Detection of Bacterial Outer Membrane Vesicles. Cell Reports. https://doi.org/10.1016/j.celrep.2019.01.062

6. Nakatogawa, H., Suzuki, K., Kamada, Y., & Ohsumi, Y. (2009). Dynamics and diversity in autophagy mechanisms: Lessons from yeast. Nature Reviews Molecular Cell Biology. https://doi.org/10.1038/nrm2708

7. Tsukada, M., & Ohsumi, Y. (1993). Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae. FEBS Letters. https://doi.org/10.1016/0014-5793(93)80398-E

8. Wild, P., McEwan, D. G., & Dikic, I. (2014). The LC3 interactome at a glance. Journal of Cell Science. https://doi.org/10.1242/jcs.140426

9. Igloi, G. L. (2001). Cloning, expression patterns, and chromosome localization of three human and two mouse homologues of GABAA receptor-associated protein. Genomics. https://doi.org/10.1006/geno.2001.6555

10. Glick, D., Barth, S., & Macleod, K. F. (2010). Autophagy: Cellular and molecular mechanisms. Journal of Pathology. https://doi.org/10.1002/path.2697

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, Ch, Gp, Ha, In, Pm, Pm, Rb, SyHa, Ze
Applications: WB, Simple Western, ELISA, Flow, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, PLA, ChIP, KD, KO

Publications for LC3B Antibody (NB100-2220)(1098)

We have publications tested in 18 confirmed species: Human, Mouse, Rat, Alligator, Avian, Bacteria, Bovine, Canine, Chicken, E. coli, Golden Syrian Hamster, Guinea Pig, Hamster, Invertebrate, Monkey, Porcine, Primate, Zebrafish.

We have publications tested in 14 applications: ChIP, ELISA, EM, FLOW, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KD, KO, PLA, WB.


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(162)
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(44)
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(11)
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(25)
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(7)
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(4)
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(4)
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(1)
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Showing Publications 1 - 10 of 1098. Show All 1098 Publications.
Publications using NB100-2220 Applications Species
de Carvalho NZM, Chiarotto GB, Bernardes D et al. Neuroprotection by dimethyl fumarate following ventral root crush in C57BL/6 J mice Brain Res. Bull. Aug 28 2020 [PMID: 32866558] (WB) WB
Gao Y, Zhang W, Zeng L et al. Exercise and dietary intervention ameliorate high-fat diet-induced NAFLD and liver aging by inducing lipophagy Redox Biology Jan 1 2020 [PMID: 32863214]
Xu M, Hu Y, Ding W et al. Rationally designed rapamycin-encapsulated ZIF-8 nanosystem for overcoming chemotherapy resistance Biomaterials Aug 8 2020 [PMID: 32841911] (WB, Human) WB Human
Mukhopadhyay S, Praharaj PP, Naik PP et al. Identification of Annexin A2 as a key mTOR target to induce autophagy to autophagy dependent cell death in stress Biochim Biophys Acta Mol Basis Dis Aug 22 2020 [PMID: 32841734] (WB) WB
Lauterbach MA, Saavedra V, Mangan MSJ et al. 1-Deoxysphingolipids cause autophagosome and lysosome accumulation and trigger NLRP3 inflammasome activation Autophagy Aug 24 2020 [PMID: 32835606] (WB, Mouse) WB Mouse
Thomas A, Samykutty A, Gomez-Gutierrez JG et al. Actively Targeted Nanodelivery of Echinomycin Induces Autophagy-Mediated Death in Chemoresistant Pancreatic Cancer In Vivo Cancers (Basel) Aug 14 2020 [PMID: 32823919] (WB, Human) WB Human
Li C, Zhao W, Pan X et al. LncRNA KTN1-AS1 promotes the progression of non-small cell lung cancer via sponging of miR-130a-5p and activation of PDPK1 Oncogene Aug 20 2020 [PMID: 32820252] (WB, Human) WB Human
McCauley ME, O'Rourke JG, Yanez A et al. C9orf72 in myeloid cells suppresses STING-induced inflammation Nature Sep 1 2020 [PMID: 32814898] (WB, Mouse) WB Mouse
Park HJ, Jo DS, Choi DS et al. Ursolic acid inhibits pigmentation by increasing melanosomal autophagy in B16F1 cells Biochem. Biophys. Res. Commun. Aug 10 2020 [PMID: 32792197] (WB, Mouse) WB Mouse
Abokyi S, Shan S, To C et al. Autophagy Upregulation by the TFEB Inducer Trehalose Protects against Oxidative Damage and Cell Death Associated with NRF2 Inhibition in Human RPE Cells Oxidative Medicine and Cellular Longevity Jul 22 2020 [PMID: 32774677] (WB, Human) WB Human
Show All 1098 Publications.

Reviews for LC3B Antibody (NB100-2220) (47) 4.747

Average Rating: 4.7
(Based on 47 reviews)
We have 47 reviews tested in 7 species: Human, Mouse, Rat, Mouse and Human, Mouse and insect, Other, murine.

Reviews using NB100-2220:
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WB
(37)
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(2)
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(3)
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(1)
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(1)
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(1)
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(1)
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(19)
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(17)
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(4)
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(2)
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Images Ratings Applications Species Date Details
Western Blot LC3B NB100-2220
Enlarge
reviewed by:
Hongxue Shi
WB Mouse 10/02/2020
View

Summary

ApplicationWestern Blot
Sample Testedmouse liver lysate
SpeciesMouse
LotEP
Western Blot LC3B NB100-2220
Enlarge
reviewed by:
Anonymous
WB Mouse 09/15/2020
View

Summary

ApplicationWestern Blot
Sample TestedLiver tissue
SpeciesMouse
LotDC