LC3B Antibody (1251A)

Images

 
Genetic Strategies: Knockout Validated: LC3B Antibody (1251A) [NBP2-46892] - Western Blot analysis shows lysates of HeLa human cervical epithelial carcinoma parental cell line and LC3B knockout HeLa cell line ...read more
Genetic Strategies: Knockout Validated: LC3B Antibody (1251A) [NBP2-46892] - LC3B was detected in immersion fixed Chloroquine treated Hela cells (left) but was not detected in LC3B knockout Hela cells (right) ...read more
Western Blot: LC3B Antibody (1251A) [NBP2-46892] - The expression of LC3B in rat tissue. This image was submitted via customer Review.
Biological Strategies: Flow (Intracellular): LC3B Antibody (1251A) [NBP2-46892] - Jurkat cells were either untreated (A) or treated with 50uM chloroquine for 24 hours (B). An intracellular stain was performed ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody (1251A) [NBP2-46892] - HeLa cells were treated with Chlorquine for 24 hours prior to fixation, permeabilization and incubation with anti- [Catalog # NBP2-46892] and ...read more
Immunohistochemistry-Paraffin: LC3B Antibody (1251A) [NBP2-46892] - IHC (Immunohistochemical) analysis of a formalin fixed and paraffin embedded tissue section of normal mouse brain using rabbit monoclonal LC3B (1251A) ...read more
Flow Cytometry: LC3B Antibody (1251A) [NBP2-46892] - HeLa cells untreated or treated with 50 uM chloroquine for 24h. Image from the Alexa Fluor 405 version of this antibody.
Biological Strategies: Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Analysis shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with Chloroquine. PVDF membrane ...read more
Biological Strategies: Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Western Blot analysis using HeLa cells treated with (+) or without (-) Chloriquine (CQ). A specific band was detected for LC3B at a ...read more
Biological Strategies: Western Blot: LC3B Antibody (1251A) [NBP2-46892] - Western Blot image of monoclonal anti-LC3B Antibody (Clone 1251D) [Catalog # NBP2-46892]. HeLa and Neuro2A cells were treated with or ...read more
Immunocytochemistry/ Immunofluorescence: LC3B Antibody (1251A) [NBP2-46892] - HeLa cells were treated with 50 uM Chloroquine for 24 hour prior to fixation in 10% buffered formalin for 10 min. Cells were permeabilized in ...read more
Flow (Intracellular): LC3B Antibody (1251A) [NBP2-46892] - HeLa human cervical epithelial carcimoa cell line either treated with 50 uM chloroquine for 24 hours (filled histogram) or untreated (open histogram) was ...read more
Immunoprecipitation: LC3B Antibody (1251A) [NBP2-46892] - Western blot analysis of LC3 immunoprecipitation. 10 ug of monoclonal anti-LC3B antibody (clone 1251A) [Catalog # NBP2-46892] was used to immunoprecipitate LC3 ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Rt, Po, Bt, Bv, Fi, Xp, ZeSpecies Glossary
Applications WB, Flow, Flow-IC, ICC/IF, IHC, IHC-P, IP, KO
Clone
1251A
Clonality
Monoclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml

Order Details

LC3B Antibody (1251A) Summary

Additional Information
Recombinant Monoclonal Antibody.
Immunogen
Recombinant monoclonal LC3B Antibody (1251A) 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
Autophagosomes
Predicted Species
Rat (94%), Porcine (100%), Bovine (100%), Fish (100%), Zebrafish (100%), Bat (100%), Xenopus (94%). Backed by our 100% Guarantee.
Isotype
IgG
Clonality
Monoclonal
Host
Rabbit
Gene
MAP1LC3B
Purity
Protein G 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
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 10-20 ug/ml
  • Immunohistochemistry 1:100-1:500
  • Immunohistochemistry-Paraffin 1:100-1:500
  • Immunoprecipitation 2-10 ug
  • Flow (Intracellular) 1 - 2.5 ug/mL
  • Knockout Validated
Application Notes
In WB this LC3B recombinant monoclonal antibody detects both LC3B I and LC3B II with chloroquine treatment. With ICC autophagosome staining was observed after treatment with chloroquine.
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
LC3B Overexpression Lysate
Neuro2a Chloroquine Treated / Untreated Cell Lysate
HeLa Chloroquine Treated / Untreated Cell Lysate
Reviewed Applications
Read 3 Reviews rated 5
using
NBP2-46892 in the following applications:

Publications
Read Publications using
NBP2-46892 in the following applications:

Reactivity Notes

Mouse and Rat reactivity reported from verified customer reviews.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Alternate Names for LC3B Antibody (1251A)

  • Apg8b
  • ATG8F
  • Autophagy-related protein LC3 B
  • Autophagy-related ubiquitin-like modifier LC3 B
  • lc3b autophagy marker
  • LC3B monoclonal
  • 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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NBP2-46892
Species: Hu, Mu, Rt, Rt, Po, Bt, Bv, Fi, Xp, Ze
Applications: WB, Flow, Flow-IC, ICC/IF, IHC, IHC-P, IP, KO

Publications for LC3B Antibody (NBP2-46892)(4)

Reviews for LC3B Antibody (NBP2-46892) (3) 53

Average Rating: 5
(Based on 3 reviews)
We have 3 reviews tested in 3 species: Human, Mouse, Rat.

Reviews using NBP2-46892:
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WB
(3)
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Human
(1)
Mouse
(1)
Rat
(1)
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Images Ratings Applications Species Date Details
Western Blot LC3B NBP2-46892
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reviewed by:
Anonymous
WB Rat 07/29/2017
View

Summary

ApplicationWestern Blot
Sample Tested Rat tissue
SpeciesRat
Western Blot LC3B NBP2-46892
Enlarge
reviewed by:
Anonymous
WB Mouse 09/28/2016
View

Summary

ApplicationWestern Blot
Sample TestedLiver
SpeciesMouse

Blocking

Blocking Details-

Details

Detection Notes-
Western Blot LC3B NBP2-46892
Enlarge
reviewed by:
Anonymous
WB Human 09/28/2016
View

Summary

ApplicationWestern Blot
Sample Testedbladder cell line
SpeciesHuman

Blocking

Blocking Details-

Details

Detection Notes-

Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for LC3B Antibody (NBP2-46892). (Showing 1 - 2 of 2 FAQs).

  1. Hello, somewhere on the LC3B antibody page I read it is recommended to use a 0.2uM membrane for western blot?
    • Yes, because LC3B-I is about 14-15 kDa and LC3B-II is even even smaller, the protein may slip through the 0.45 uM membrane and be lost during blotting. However, there has been success using 0.45 uM membranes; a 0.2 uM membrane is something to consider if no signal is detected.
  2. What should be the composition of the lysis buffer? How much protein should be loaded per lane? 
    • We load 10 ug of total protein and the lysates are prepared in RIPA buffer. In addition, the gel must be 15-20% to maximize the resolution of LC3 and the membrane should be 0.2um PVDF.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NBP2-46892AF350
Alexa Fluor 405 NBP2-46892AF405
Alexa Fluor 488 NBP2-46892AF488
Alexa Fluor 532 NBP2-46892AF532
Alexa Fluor 594 NBP2-46892AF594
Alexa Fluor 647 NBP2-46892AF647
Alexa Fluor 700 NBP2-46892AF700
Alexa Fluor 750 NBP2-46892AF750
Allophycocyanin NBP2-46892APC
Biotin NBP2-46892B
DyLight 350 NBP2-46892UV
DyLight 405 NBP2-46892V
DyLight 488 NBP2-46892G
DyLight 550 NBP2-46892R
DyLight 594 NBP2-46892DL594
DyLight 650 NBP2-46892C
DyLight 680 NBP2-46892FR
DyLight 755 NBP2-46892IR
FITC NBP2-46892F
HRP NBP2-46892H
Janelia Fluor 549 NBP2-46892JF549
Janelia Fluor 646 NBP2-46892JF646
PE NBP2-46892PE
PerCP NBP2-46892PCP

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Recent Reviews

5
5
3
4
0
3
0
2
0
1
0

Anonymous
07/29/2017
Application: WB
Species: Rat

Anonymous
09/28/2016
Application: WB
Species: Mouse

Anonymous
09/28/2016
Application: WB
Species: Human

Bioinformatics

Gene Symbol MAP1LC3B
Entrez
Uniprot