LC3B Antibody [Alexa Fluor® 647]

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Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Ba, Bv, Ca, Pm, ZeSpecies Glossary
Applications WB, Flow, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IHC-FrFl, KD, KO
Clonality
Polyclonal
Host
Rabbit
Conjugate
Alexa Fluor 647

LC3B Antibody [Alexa Fluor® 647] Summary

Immunogen
Polyclonal LC3B Antibody was made to 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
  • Flow Cytometry
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin
  • Immunohistochemistry Free-Floating
  • Knockdown Validated
  • Knockout Validated
Application Notes
Optimal dilution of this antibody should be experimentally determined.
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.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID:32802192). 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).. Rat reactivity reported in scientific literature (30067379).. Bacteria reactivity reported in scientific literature (31110360).. Bovine reactivity reported in scientific literature (21868124).. Other species have not been tested.

Packaging, Storage & Formulations

Storage
Store at 4C in the dark.
Buffer
50mM Sodium Borate
Preservative
0.05% Sodium Azide
Purity
Immunogen affinity purified

Notes

Alexa Fluor (R) products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

Alternate Names for LC3B Antibody [Alexa Fluor® 647]

  • 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
  • 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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Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for LC3B Antibody (NB600-1384AF647). (Showing 1 - 1 of 1 FAQ).

  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.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB600-1384AF350
Alexa Fluor 405 NB600-1384AF405
Alexa Fluor 488 NB600-1384AF488
Alexa Fluor 532 NB600-1384AF532
Alexa Fluor 594 NB600-1384AF594
Alexa Fluor 647 NB600-1384AF647
Alexa Fluor 700 NB600-1384AF700
Alexa Fluor 750 NB600-1384AF750
Biotin NB600-1384B
DyLight 350 NB600-1384UV
DyLight 405 NB600-1384V
DyLight 488 NB600-1384G
DyLight 550 NB600-1384R
DyLight 594 NB600-1384DL594
DyLight 650 NB600-1384C
DyLight 680 NB600-1384FR
DyLight 755 NB600-1384IR
FITC NB600-1384F
HRP NB600-1384H
Janelia Fluor 549 NB600-1384JF549
Janelia Fluor 646 NB600-1384JF646
PE NB600-1384PE

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Array NB600-1384AF647

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Bioinformatics

Gene Symbol MAP1LC3B