LC3A Antibody

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Western Blot: LC3A Antibody [NBP1-19167] - Total protein from HeLa and Neuro2A cells treated with or without 50 uM chloroquine for 24 hours was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane ...read more
Immunocytochemistry/ Immunofluorescence: LC3A Antibody [NBP1-19167] - LC3/MAP1 [NBP1-19167] - LC3 antibody was tested in HeLa cells with Dylight 488 (green). Cells were treated overnight with 50uM chloroquine to induce ...read more
Immunohistochemistry-Paraffin: LC3A Antibody [NBP1-19167] - LC3/MAP1 [NBP1-19167] - IHC analysis of a formalin fixed and paraffin embedded tissue section of mouse brain using LC3 antibody at 1:300 dilution. The signal ...read more
Flow Cytometry: LC3/MAP1LC3A Antibody [NBP1-19167] - Staining of NTERA-2 cells using NBP1-19167 at a 1:50 dilution detected using Dylight-488 conjugated goat anti-rabbit IgG secondary antibody.
Western Blot: LC3/MAP1LC3A Antibody [NBP1-19167] - Human brain lysate.
Immunohistochemistry-Paraffin: LC3/MAP1LC3A Antibody [NBP1-19167] - IHC analysis of a formalin fixed and paraffin embedded tissue section of mouse liver using LC3 antibody at 1:300 dilution. The signal was developed ...read more
Simple Western: LC3/MAP1LC3A Antibody [NBP1-19167] - 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, Bv, ZeSpecies Glossary
Applications WB, Simple Western, Flow, ICC/IF, IHC, IHC-Fr, IHC-P
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
0.2 mg/ml
   

Biological Strategies

     

Order Details

View Available Conjugates
Catalog# & Conjugate Size Price

LC3A Antibody Summary

Immunogen
Genomic sequence made to an N-terminal portion of the human LC3A protein [Swiss-Prot# Q9H492].
Localization
Autophagosomes. Cytoplasm. Intracytoplasmic membrane; Lipid-anchor. Cytoplasmic vesicle autophagosome membrane; Lipid-anchor. Note= LC3-II binds to the autophagic membranes.
Marker
Autophagosome Marker
Specificity
Although specificity between LC3A and LC3B has not been tested, this antibody was created to a peptide that has 100% identity to LC3A and 62% identity to LC3B.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
MAP1LC3A
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 1:2500
  • Simple Western 1:40
  • Flow Cytometry 1:100
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:100 - 1:400
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:100 - 1:400
Application Notes
In WB, bands are seen at approx. 14-17 kDa position. In ICC/IF, autophagosome formation has been seen in HeLa cells after treatment with 50uM chloroquine. Use in Immunohistochemistry-Frozen reported in scientific literature (PMID: 23936035).

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.
Theoretical MW
14 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.
Positive Control
LC3A Lysate (NBL1-12843)
Brain Lysate (NB820-59177)
Brain Thalamus Lysate (NB820-59199)
Neuro2a Lysate (NBP2-49688)
HeLa Lysate (NBP2-49689)
Reviewed Applications
Read 2 Reviews rated 4.5
using
NBP1-19167 in the following applications:

Publications
Read Publications using
NBP1-19167 in the following applications:

Reactivity Notes

Human, mouse, bovine, rat and zebrafish.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS, 0.1% BSA, and 50% Glycerol
Preservative
0.05% Sodium Azide
Concentration
0.2 mg/ml
Purity
Immunogen affinity purified

Alternate Names for LC3A Antibody

  • Apg8
  • APG8a
  • Apg8p3
  • ATG8E
  • Autophagy-related protein LC3 A
  • Autophagy-related ubiquitin-like modifier LC3 A
  • LC3
  • LC3A
  • lc3-i, ii
  • MAP1A/1B light chain 3 A
  • MAP1ALC3
  • MAP1ALC3MAP1A/MAP1B LC3 A
  • MAP1BLC3MAP1A/MAP1B light chain 3 A
  • MAP1LC3A
  • microtubule-associated protein 1 light chain 3 alphaMAP1 light chain 3-like protein 1
  • microtubule-associated proteins 1A/1B light chain 3
  • microtubule-associated proteins 1A/1B light chain 3A
  • MLP3A

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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NBP1-19167
Species: Hu, Mu, Rt, Bv, Ze
Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-Fr, IHC-P

Publications for LC3A Antibody (NBP1-19167)(16)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 5 applications: ICC/IF, IHC, IHC-Fr, IHC-P, WB.


Filter By Application
ICC/IF
(1)
IHC
(1)
IHC-Fr
(1)
IHC-P
(3)
WB
(9)
All Applications
Filter By Species
Human
(9)
Mouse
(6)
All Species
Showing Publications 1 - 10 of 16. Show All 16 Publications.
Publications using NBP1-19167 Applications Species
Kohler LJ, Reed SR, Sarraf SA et al. Effector Protein Cig2 Decreases Host Tolerance of Infection by Directing Constitutive Fusion of Autophagosomes with the Coxiella-Containing Vacuole MBio 2016 Jul 20 [PMID: 27435465] (WB). Elife. 2016 Jul 20 [PMID: 27435465] (WB)

Details:
This citation used the PE version of this antibody.
WB
Saito T, Asai K, Sato S et al. Autophagic vacuoles in cardiomyocytes of dilated cardiomyopathy with initially decompensated heart failure predict improved prognosis Autophagy Feb 18 2016 [PMID: 26890610] (IHC-P, Human) IHC-P Human
Campos T, Ziehe J, Palma M et al. Rheb promotes cancer cell survival through p27Kip1-dependent activation of autophagy Mol. Carcinog. 2015 Jan 15 [PMID: 25594310] (WB, Human) WB Human
Yang Annan, Rajeshkumar N V, Wang Xiaoxu et al. Autophagy is critical for pancreatic tumor growth and progression in tumors with p53 alterations. Cancer Discov. 2014 May 29 [PMID: 24875860]
Rao S, Tortola L, Perlot T et al. A dual role for autophagy in a murine model of lung cancer. Nat Commun. 2014 [PMID: 24445999] (IHC-P, Mouse) IHC-P Mouse
Giovannini M, Bonne NX, Vitte J et al. mTORC1 inhibition delays growth of neurofibromatosis type 2 schwannoma. Neuro-oncology 2014 Jan 10 [PMID: 24414536] (IHC-P, Mouse) IHC-P Mouse
Bruntz RC, Taylor HE, Lindsley CW, Brown HA. Phospholipase D2 mediates survival signaling through direct regulation of Akt in glioblastoma cells. J Biol Chem. 2013 Nov 20 [PMID: 24257753] (WB, Human) WB Human
Howell GM, Gomez H, Collage RD et al. Augmenting Autophagy to Treat Acute Kidney Injury during Endotoxemia in Mice. PLoS One 2013 Jul 30 [PMID: 23936035] (IHC-Fr, ICC/IF, Mouse) IHC-Fr, ICC/IF Mouse
Santoni M, Amantini C, Morelli MB et al. Pazopanib and sunitinib trigger autophagic and non-autophagic death of bladder tumour cells. Br J Cancer 2013 Jul 25 [PMID: 23887605] (WB, Human) WB Human
Kuo SH, Tang G, Ma K et al. Macroautophagy abnormality in essential tremor PLoS One 2012 [PMID: 23300858] (WB, Human) WB Human
Show All 16 Publications.

Reviews for LC3A Antibody (NBP1-19167) (2) 4.52

Average Rating: 4.5
(Based on 2 reviews)
We have 2 reviews tested in 2 species: Mouse, Rat.
We have 2 reviews tested in 2 application: ICC, WB.

Reviews using NBP1-19167:
Filter by Applications
ICC
(1)
WB
(1)
All Applications
Filter by Species
Mouse
(1)
Rat
(1)
All Species
Images Ratings Applications Species Date Details
Immunocytochemistry LC3A NBP1-19167
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reviewed by:
Tania Campos
ICC Mouse 06/12/2015
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Summary

ApplicationImmunocytochemistry
Sample TestedMEF cells
SpeciesMouse
LotC-1

Blocking

Blocking Details1% BSA

Primary Anitbody

Dilution RatioLC3/MAP1LC3A Antibody (NBP1-19167) 1:500 on PBS 1% BSA (8 h)

Secondary Antibody

Secondary Descriptionanti-rabbit IgG Alexa 546 (Invitrogen)
Secondary Manufacturer Cat#A11010
Secondary Concentration1:1000

Details

Detection NotesConfocal microscope
Fixation DetailsFixed: 4% PFA (15 min), Permeabilization: 0.1% Triton X-100 on PBS 1% BSA (10 min)
Wash DescriptionPBS pH 7.4
Western Blot LC3A NBP1-19167
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reviewed by:
Anonymous
WB Rat 02/24/2015
View

Summary

ApplicationWestern Blot
Sample TestedRat bone marrow stromal cells
SpeciesRat

Blocking

Blocking DetailsBSA, 5%, 1 hour, RT

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG

Details

Detection NotesECL

Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for LC3A Antibody (NBP1-19167). (Showing 1 - 3 of 3 FAQs).

  1. May we ask if it is possible to perform IF to stain LC3-I and LC3-II separately with two different fluorescent colors?
    • Yes, it is possible to perform IF stain for LC3-I and LC3-II separately with two different fluorescent colors! You will have to use two different primary antibodies and in order to avoid any potential background/cross reactivity issues, I would suggest that you employ conjugated primary antibodies for the testing. 1. Our LC3I antibody (NBP1-78964) has been designed to specifically detect the cytosolic form of the LC3 protein which is actually LC3 I (Note: LC3-II binds to the autophagic membranes). 2. There is not even a single antibody to our knowledge that would exclusively detect the LC3 II form, and you would have to detect LC3II/ autophagic membranes form with an antibody which detects LC3 I/LC3II together. Therefore you may opt second antibody from one of the followings: LC3 Antibody (NB100-2220), LC3 Antibody (NB100-2331), LC3 Antibody (NBP1-19167). All of these mentioned catalog #s come with different options for their conjugated forms and you may select appropriate conjugated forms for performing the IF staining using our explained criteria.
  2. We have received an antibody against LC3 (# NBP1-19167) and I need a precision regarding its specificity. According to the datasheet, the antibody was raised against a peptide 100% identical to LC3A and 62% to LC3B. My question is: Is LC3A used as a synonym for LC3-I and LC3B as a synonym of LC3-II?
    • LC3A is not a synonym for LC3-I or LC3-II. When we refer to LC3-I and -II, we actually mean LC3B-I and LC3B-II. I hope this helps, but please let me know if you have any questions.
  3. We purchased an antibody against LC3 (NBP1-19167) from your company. According to the information found on your web site, three papers used this antibody to detect both the LC3-I and LC3-II form of the protein. The catalog number of the antibody is not mentioned in the papers. How can I be sure that it is the one used by the authors? It is important since I want to know if this antibody NBP1-19167 recognize both LC3 forms.
    • Our publication evaluation team is highly qualified and we found that these publications were cited under # NBP1-19167 after consulting the archived order information of the customers. As far as the detection of the two forms is concerned, any given LC3 antibody should detect LC3-I as well as LC3-II forms because the only difference between the two forms is that LC3-II carries the phosphatidylethanolamine (PE) conjugate whereas LC3-I does not have a PE conjugate. In nutshell, following translation, the unprocessed form of LC3 (pro-LC3) is proteolytically cleaved by Atg4 protease, resulting in the cytosolic form of LC3 (LC3-I) with a carboxyterminal exposed glycine. Upon induction of autophagy, the 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 the highly lipophilic PE moiety to generate LC3-II (which is recruited to autophagosomal membranes). Therefore, the presence of both bands (LC3-I & LC3-II) will depend upon the extent of autophagy in your particular samples.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

PE Labeled NBP1-19167PE

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Recent Reviews

4.5
5
1
4
1
3
0
2
0
1
0

Tania Campos
06/12/2015
Application: ICC
Species: Mouse

Anonymous
02/24/2015
Application: WB
Species: Rat

Bioinformatics

Gene Symbol MAP1LC3A
Entrez
Uniprot