LC3B Antibody

Images

 
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB600-1384] - LC3B/MAP1 [NB600-1384] - IF Confocal analysis of HeLa cells using LC3B antibody (NB600-1384, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB600-1384] - LC3B/MAP1 [NB600-1384] - LC3 antibody was tested in 50uM Chloroquine treated HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB600-1384] - LC3B/MAP1 [NB600-1384] - Analysis using the HRP conjugate of NB600-1384. Staining of treated U373-MG cells using NB600-1384. The nuclei were stained ...read more
Western Blot: LC3B Antibody [NB600-1384] - LC3B/MAP1 [NB600-1384] - analysis of LC3B in IB3-1 whole cell lysate using anti-LC3B antibody. Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB600-1384] - LC3B/MAP1 [NB600-1384] - analysis of LC3B in HeLa cells using anti-LC3B antibody (red). Nuclei were counterstained with DAPI (blue).
Western Blot: LC3B Antibody [NB600-1384] - LC3B/MAP1 [NB600-1384] - Detection of LC3B in treated U87-MG (human glioblastoma astrocytoma) lysates.
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB600-1384] - 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 [NB600-1384] - LC3B/MAP1 [NB600-1384] - Staining of treated U373-MG (human glioblastoma.
Flow Cytometry: LC3B/MAP1LC3B Antibody [NB600-1384] - Staining of NTERA-2 cells using NB600-1384 at a 1:200 dilution detected using Dylight-488 conjugated goat anti-rabbit IgG secondary.
Immunocytochemistry/ Immunofluorescence: LC3B Antibody [NB600-1384] - 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
Immunohistochemistry-Paraffin: LC3B/MAP1LC3B Antibody [NB600-1384] - analysis of U87MG glioma xenografts using anti-LC3B antibdoy. Image from verified customer review.
Immunohistochemistry: LC3B/MAP1LC3B Antibody [NB600-1384] - LC3B staining in treated U87-MG cultured & subcutaneous tumors.
Immunohistochemistry: LC3B/MAP1LC3B Antibody [NB600-1384] - LC3B staining in gliobastoma multiform tissue.
Simple Western: LC3B/MAP1LC3B Antibody [NB600-1384] - Simple Western lane view shows a specific band for LC3B 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, Bv, Ca, Pm, ZeSpecies Glossary
Applications WB, Simple Western, EM, Flow, ICC/IF, IHC, IHC-Fr, IHC-P
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml
   

Biological Strategies

     

LC3B Antibody Summary

Immunogen
A synthetic peptide made to the N-terminal region of the human LC3B protein. [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 1:1000
  • Simple Western 1:100
  • Electron Microscopy
  • Flow Cytometry 1:200
  • Immunocytochemistry/Immunofluorescence 0.1-2 ug/ml
  • Immunohistochemistry
  • Immunohistochemistry-Frozen 1:400
  • Immunohistochemistry-Paraffin 1:200-1:400
Application Notes
This LC3 antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry-frozen sections, Flow Cytometry and Western Blot where bands are seen ~17 and 19 kDa corresponding to LC3-II and LC3-I. Electron Microscopy was reported in scientific literature.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Positive Control
Brain Hippocampus Lysate (NB820-60577)
Brain Lysate (NB820-59177)
Neuro2a Lysate (NBP1-78158)
Neuro2a Lysate (NBP2-49688)
HeLa Lysate (NBP2-49689)
Reviewed Applications
Read 15 Reviews rated 4.2
using
NB600-1384 in the following applications:

Publications
Read Publications using
NB600-1384 in the following applications:

Reactivity Notes

Zebrafish reactivity reported in scientific literature (PMID: 23724125). Canine and primate reactivity reported in scientific literature (PMID: 24027311) Porcine reactivity reported in scientific literature (PMID: 25378587) Other species have not been tested.

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

  • Apg8b
  • ATG8F
  • Autophagy-related protein LC3 B
  • Autophagy-related ubiquitin-like modifier LC3 B
  • lc3b autophagy marker
  • 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

Autophagy is a process of intracellular bulk degradation in which cytoplasmic components, including organelles, are sequestered within double-membrane vesicles that deliver the contents to the lysosome/vacuole for degradation. During macroautophagy, the sequestering vesicles, termed autophagosomes, fuse with the lysosome or vacuole resulting in the delivery of an inner vesicle (autophagic body) into the lumen of the degradative compartment. There are 16 proteins participating in the autophagy pathway in humans. The autophagy protein LC3, a mammalian homologue of Atg8, was originally identified as microtubule-associated protein 1 light chain 3. It is a component of both the MAP1A and MAP1B microtubule-binding domains and the heavy-chain independent regulation of LC3 expression may modify MAP1 microtubule-binding activity during development. LC3 is the only known mammalian protein identified that stably associates with the autophagosome membranes. LC3-I is cytosolic and LC3-II is membrane bound and enriched in the autophagic vacuole fraction. The detection of LC3-I to LC3-II conversion is a useful and sensitive marker for distinguishing autophagy in mammalian cells.

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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NB600-1335
Species: Hu, Mu, Rt, Pm
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NB100-56098
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NBP2-57604
Species: Hu
Applications: ICC/IF
NB600-1384
Species: Hu, Mu, Rt, Po, Bv, Ca, Pm, Ze
Applications: WB, Simple Western, EM, Flow, ICC/IF, IHC, IHC-Fr, IHC-P

Publications for LC3B Antibody (NB600-1384)(189)

We have publications tested in 8 confirmed species: Human, Mouse, Rat, Bovine, Canine, Porcine, Primate, Zebrafish.

We have publications tested in 9 applications: EM, FLOW, ICC/IF, IHC, IHC-Fr, IHC-FrFl, IHC-P, IP, WB.


Filter By Application
EM
(2)
FLOW
(6)
ICC/IF
(36)
IHC
(9)
IHC-Fr
(2)
IHC-FrFl
(1)
IHC-P
(7)
IP
(1)
WB
(117)
All Applications
Filter By Species
Human
(78)
Mouse
(40)
Rat
(10)
Bovine
(1)
Canine
(2)
Porcine
(2)
Primate
(1)
Zebrafish
(2)
All Species
Showing Publications 1 - 10 of 189. Show All 189 Publications.
Publications using NB600-1384 Applications Species
Parcon PA, Balasubramaniam M, Ayyadevara S et al. Apolipoprotein E4 inhibits autophagy gene products through direct, specific binding to CLEAR motifs Alzheimers Dement 2017 Sep 22 [PMID: 28945989] (Human) Human
Kemp V, Dautzenberg IJC, Limpens RW et al. Oncolytic Reovirus Infection Is Facilitated by the Autophagic Machinery Viruses 2017 Sep 21 [PMID: 28934149] (WB, Human) WB Human
Muniz-Feliciano L, Doggett TA, Zhou Z, Ferguson TA. RUBCN/rubicon and EGFR regulate lysosomal degradative processes in the retinal pigment epithelium (RPE) of the eye Autophagy 2017 Sep 21 [PMID: 28933590] (WB) WB
Niklaus M, Adams O, Berezowska S et al. Expression analysis of LC3B and p62 indicates intact activated autophagy is associated with an unfavorable prognosis in colon cancer Oncotarget 2017 Aug 15 [PMID: 28903368] (IHC-P, Human) IHC-P Human
Jiang P, Gan M, Yen SH et al. Impaired endo-lysosomal membrane integrity accelerates the seeding progression of a-synuclein aggregates Sci Rep 2017 Aug 09 [PMID: 28794446] (ICC/IF, Human) ICC/IF Human
Wiedmer T, Blank A, Pantasis S et al. Autophagy inhibition improves sunitinib efficacy in pancreatic neuroendocrine tumors via a lysosome-dependent mechanism Mol. Cancer Ther. 2017 Jul 20 [PMID: 28729403] (Human) Human
Germic N, Stojkov D, Oberson K, et al. Neither eosinophils nor neutrophils require ATG5-dependent autophagy for extracellular DNA trap formation Immunology 2017 Jul 13 [PMID: 28703297] (Mouse) Mouse
Cao B, Parnell LA, Diamond MS, Mysorekar IU. Inhibition of autophagy limits vertical transmission of Zika virus in pregnant mice J. Exp. Med. 2017 Jul 10 [PMID: 28694387] (WB, Mouse) WB Mouse
Sun QZ, Lin GF, Li LL et al. Discovery of Potent and Selective Inhibitors of Cdc2-Like Kinase 1 (CLK1) as a New Class of Autophagy Inducers J. Med. Chem. 2017 Jul 27 [PMID: 28692292] (WB) WB
Ran G, Ying L, Li L et al. Resveratrol ameliorates diet-induced dysregulation of lipid metabolism in zebrafish (Danio rerio) PLoS ONE 2017 Jul 07 [PMID: 28686680] (WB, Zebrafish) WB Zebrafish
Show All 189 Publications.

Reviews for LC3B Antibody (NB600-1384) (15) 4.215

Average Rating: 4.2
(Based on 15 reviews)
We have 15 reviews tested in 4 species: Human, Mouse, Rat, Other.
We have 15 reviews tested in 4 application: WB, IF, ICC, IHC.

Reviews using NB600-1384:
Filter by Applications
WB
(9)
IF
(3)
ICC
(1)
IHC
(1)
All Applications
Filter by Species
Human
(6)
Mouse
(6)
Rat
(2)
Other
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot LC3B NB600-1384
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reviewed by:
Anonymous
WB Mouse 07/22/2016
View

Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate
SpeciesMouse
LotBQ

Blocking

Blocking Details5% skim milk in TBS

Primary Anitbody

Dilution Ratio1:1,000

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG, HRP
Secondary Concentration0.4 ug/ml

Details

Detection NotesECL, exposure 30 seconds
Western Blot LC3B NB600-1384
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reviewed by:
Anonymous
WB Mouse 06/28/2016
View

Summary

ApplicationWestern Blot
Sample Testedmice primary macrophages
SpeciesMouse
LotBO

Blocking

Blocking Details5% milk for 1 hour at room temperature

Primary Anitbody

Dilution Ratio4 degree overnight, 1:1,000 dilution

Secondary Antibody

Secondary Descriptiongoat anti-rabbit IgG, HRP
Secondary Concentration0.4 ug/ml

Details

Detection Noteswith ECL solution, 25 secs of exposure time.
 LC3B NB600-1384
Enlarge
reviewed by:
Anonymous
Human 02/04/2015
View

Summary

SpeciesHuman
FileView PDF
 
reviewed by:
Anonymous
IF Mouse 12/23/2014
View

Summary

ApplicationImmunofluorescence
Sample TestedSee PMID 23525089
SpeciesMouse

Blocking

Blocking DetailsSee PMID 23525089

Details

Detection NotesSee PMID 23525089
Fixation DetailsMethanol fixation required.

Comments

CommentsIndependent review from antibodyresource.comPublished in PMID 23525089.
 
reviewed by:
Anonymous
IF Rat 12/23/2014
View

Summary

ApplicationImmunofluorescence
Sample TestedSee PMID 23688351
SpeciesRat

Blocking

Blocking DetailsSee PMID 23688351

Details

Detection NotesSee PMID 23688351

Comments

CommentsIndependent review from antibodyresource.comPublished in PMID 23688351.
Western Blot LC3B NB600-1384
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reviewed by:
Anonymous
WB Human 11/25/2014
View

Summary

ApplicationWestern Blot
Sample TestedIB3-1 whole cell lysate
SpeciesHuman
Lottrial sample

Blocking

Blocking Details3% non-fat dry milk (Bio-Rad) in TBST, 30 min at room temperature

Primary Anitbody

Dilution Ratio1:1000 dilution in blocking buffer, overnight at 4 degree

Secondary Antibody

Secondary Descriptionanti-rabbit IgG, HRP
Secondary Manufacturer Cat#Cell signaling, 7074S
Secondary Concentration1:5000 dilution

Details

Detection Notesfilm detection, using Clarity Western ECL (Bio-Rad) for 3 min.
Western Blot LC3B NB600-1384
Enlarge
reviewed by:
Anonymous
WB Mouse 11/23/2014
View

Summary

ApplicationWestern Blot
Sample Testedmouse hearts
SpeciesMouse

Blocking

Blocking DetailsBSA

Primary Anitbody

Dilution Ratio1:2000

Secondary Antibody

Secondary Descriptionrabbit anti-mouse IgG, HRP
Secondary Concentration1:10000

Details

Detection Notesexposure time 1 min
Western Blot LC3B NB600-1384
Enlarge
reviewed by:
Anonymous
WB Rat 11/01/2014
View

Summary

ApplicationWestern Blot
Sample TestedRat brain
SpeciesRat
CommentsWorks best with milk, detect 2 nice bands.

Blocking

Blocking DetailsMilk 10%in PBST for 2 hrs at RT

Primary Anitbody

Dilution Ratio1/2000 for 6 hrs at 4 degrees

Secondary Antibody

Secondary DescriptionDonkey anti Rabbit HRP
Secondary Manufacturer Cat#Pierce
Secondary Concentration1/10000

Details

Detection NotesECL with biorad chemidoc XRS

Comments

CommentsWorks best with milk, detect 2 nice bands.
 
reviewed by:
Anonymous
WB Mouse 10/03/2014
View

Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate alveolar macrophages
SpeciesMouse

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG HRP
Western Blot LC3B NB600-1384
Enlarge
reviewed by:
Mohammad Abedi
WB Human 07/10/2014
View

Summary

ApplicationWestern Blot
Sample TestedNeuroblastoma whole cell lysate
SpeciesHuman
Lot1384

Blocking

Blocking Details5% milk solution overnight at 4oC

Primary Anitbody

Dilution Ratio1:1000, 1 hour Incubation at RT in TBST

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG- HRP Conjugated
Secondary Manufacturer Cat##7074S
Secondary Concentration1:1000 in TBST for 1 hour at RT

Details

Detection NotesDensitometry, VersaDoc MP 5000, 300 seconds
Western Blot LC3B NB600-1384
Enlarge
reviewed by:
Anony- mous
WB Human 06/10/2014
View

Summary

ApplicationWestern Blot
SpeciesHuman
Special ApplicationsWestern blot analysis of extracts from HeLa and HT1080 cells using LC3B antibody (NB600-1384, 1:1000).
FileView PDF
Immunocytochemistry LC3B NB600-1384
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reviewed by:
Anony- mous
ICC Human 06/10/2014
View

Summary

ApplicationImmunocytochemistry
SpeciesHuman
Special ApplicationsIF Confocal analysis of HeLa cells using LC3B antibody (NB600-1384, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
FileView PDF
 
reviewed by:
Anonymous
IHC Human 10/19/2010
View

Summary

ApplicationImmunohistochemistry
Sample TestedHuman Esophagus tissue
SpeciesHuman

Details

Fixation DetailsSample Preparation: FFPE
 
reviewed by:
Anonymous
IF Other 08/28/2009
View

Summary

ApplicationImmunofluorescence
Sample TestedHuman Oral Keratinocyte
SpeciesOther
LotK
CommentsIt may be that the LC3 protein is difficult to detect in autophagosomes in Keratinocytes.

Not yet tested in HeLa.

Blocking

Blocking DetailsBlocking Buffer: BSA, Blocking Time: 1 hour, Blocking Temp: 22 degrees Celsius

Primary Anitbody

Dilution RatioPrimary Ab Incubation Dilution Buffer: 2% BSA, Primary Ab Dilution Ratio: 1/2000, Incubation Time: 16hr, Incubation Temp: 4C

Secondary Antibody

Secondary DescriptionSecondary Ab: Molecular Probes, Secondary Ab Dilution: 1/200
Secondary Manufacturer Cat#A-11034

Comments

CommentsIt may be that the LC3 protein is difficult to detect in autophagosomes in Keratinocytes.

Not yet tested in HeLa.
 
reviewed by:
Anonymous
WB Mouse 05/26/2009
View

Summary

ApplicationWestern Blot
Sample Testedmouse liver tissue, Sample Amount: 100
SpeciesMouse
LotI1

Blocking

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

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1/2000, Incubation Dilution Buffer: milk/PBS, Incubation Time: overnight, Incubation Temp: 4C

Secondary Antibody

Secondary DescriptionSecondary Ab: goat-anti rabbit-HRP (Pierce), Secondary Ab Dilution Ratio: 1/2000

Details

Detection NotesDetection Method: ECL-x-ray, Exposure Time: 1 minute, Specific Bands: 15 kDa, Non-Specific Bands: 20 kDa, 37 kDa, 82 kDa

Product General Protocols

View specific protocols for LC3B Antibody (NB600-1384): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for LC3B Antibody (NB600-1384). (Showing 1 - 4 of 4 FAQs).

  1. What is the immunogen sequence of this antibody?
    • The exact immunogen sequence is proprietary, however we can tell you that the immunogen sequence can be found between amino acids 1-50 of human LC3B.
  2. Does this antibody also recognize mouse LC3A?
    • This antibody does not cross-react with LC3A. If you are interested in detecting mouse LC3A and LC3B, you may want to consider purchasing our LC3/LC3B antibody pack, NB910-40752.
  3. Is it possible to provide me the suggested dilution of Electron Microscopy and IHC of NB600-1384?
    • Electron microscopy was performed by a customer, so unfortunately, we do not have their dilution information. For IHC, we recommend a dilution of 1:400.
  4. What % gel used for WB to obtain the separation of the LC3BI and II bands?
    • We prefer to use a 4-20% gradient gel to get ample separation between the LC3BI and II bands. If you are using a single percentage gel, a 12% or higher gel should separate the lower weights enough for proper detection.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB600-1384AF405
Alexa Fluor 488 Labeled NB600-1384AF488
Alexa Fluor 647 Labeled NB600-1384AF647
Alexa Fluor 700 Labeled NB600-1384AF700
Biotin Labeled NB600-1384B
DyLight 405 Labeled NB600-1384V
DyLight 488 Labeled NB600-1384G
DyLight 550 Labeled NB600-1384R
DyLight 650 Labeled NB600-1384C
FITC Labeled NB600-1384F
HRP Labeled NB600-1384H
PE Labeled NB600-1384PE

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Bioinformatics Tool for LC3B Antibody (NB600-1384)

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Blogs on LC3B.

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.

Breakdown: Interpreting LC3 Antibody WB Results
In rodents, MAPLC3 (Microtubule-associated protein 1 light chain 3) is expressed in the renal visceral epithelial cells, or podocytes. LC3 antibody analysis has shown the protein accumulates in its membrane-bound form of LC3II, following conversion fr...  Read full blog post.

The Role of LC3 within the Autophagic Pathway
We at Novus Biologicals have a broad antibody databasecovering the area of autophagy - over 1400 reagents in total. Autophagy is bulk degradation of cytoplasmic components - literally, self-digestion of the cell. Double-membrane vesicles, called autop...  Read full blog post.

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Recent Reviews

4.2
5
6
4
7
3
1
2
1
1
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Anonymous
07/22/2016
Application: WB
Species: Mouse

Anonymous
06/28/2016
Application: WB
Species: Mouse

Anonymous
02/04/2015
Application:
Species: Human

Bioinformatics

Gene Symbol MAP1LC3B
Entrez