Human/Mouse/Rat LC3B Antibody Summary
Accession # Q9GZQ8
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
![Detection of Human LC3B antibody by Western Blot. Detection of Human LC3B antibody by Western Blot.](https://resources.rndsystems.com/images/datasheets/antibody/LC3B_MAB85581_Western_Blot_21002.jpg)
Detection of Human LC3B by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 50 µM Chloroquine for 18 hours. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat LC3B Monoclonal Antibody (Catalog # MAB85581) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for LC3B at approximately 15 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
![Detection of Mouse and Rat LC3B antibody by Western Blot. Detection of Mouse and Rat LC3B antibody by Western Blot.](https://resources.rndsystems.com/images/datasheets/antibody/LC3B_MAB85581_Western_Blot_21003.jpg)
Detection of Mouse and Rat LC3B by Western Blot. Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line and PC-12 rat adrenal pheochromocytoma cell line untreated (-) or treated (+) with 50 µM Chloroquine for 18 hours. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat LC3B Monoclonal Antibody (Catalog # MAB85581) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for LC3B at approximately 15 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
![Detection of Mouse LC3B antibody by Simple Western<sup>TM</sup>. Detection of Mouse LC3B antibody by Simple Western<sup>TM</sup>.](https://resources.rndsystems.com/images/datasheets/antibody/LC3B_MAB85581_Simple_Western_21011.jpg)
Detection of Mouse LC3B by Simple WesternTM. Simple Western lane view shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 50 uM Chloroquine for 18 hours, loaded at 0.2 mg/mL. A specific band was detected for LC3B at approximately 17 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human LC3B Monoclonal Antibody (Catalog # MAB85581). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
![Detection of Rat LC3B antibody by Simple Western<sup>TM</sup>. Detection of Rat LC3B antibody by Simple Western<sup>TM</sup>.](https://resources.rndsystems.com/images/datasheets/antibody/LC3B_MAB85581_Simple_Western_21004.jpg)
Detection of Rat LC3B by Simple WesternTM. Simple Western lane view shows lysates of PC‑12 rat adrenal pheochromocytoma cell line untreated (-) or treated (+) with 50 uM Chloroquine for 18 hours, loaded at 0.2 mg/mL. A specific band was detected for LC3B at approximately 17 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human LC3B Monoclonal Antibody (Catalog # MAB85581). This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.
![Western Blot Show Human LC3B Antibody Specificity Using Knockout Cell Line. Western Blot Show Human LC3B Antibody Specificity Using Knockout Cell Line.](https://resources.rndsystems.com/images/datasheets/antibody/LC3B_MAB85581_Knockout_Validated_21042.jpg)
Western Blot Show Human LC3B Specificity Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 µM Chloroquine for 18 hours. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat LC3B Monoclonal Antibody (Catalog # MAB85581) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for LC3B at approximately 15 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: LC3B
Human Microtubule-associated Protein (MAP) Light Chain 3 (LC3) A is a121 amino acid (aa) protein with a predicted molecular weight of 14 kDa. It is a member of the LC3 subfamily of Autophagy-related 8 (Atg8) proteins (1). The LC3 subfamily also includes LC3B andLC3C. LC3 exhibits 100% aa sequence identity with its mouse and rat orthologs, and is orthologous to the yeast autophagy-related protein Atg8. Atg8 family members show structural similarity with Ubiquitin, but lack aa sequence similarity. LC3 was originally described as part is part of a complex that includes heavy and light chains comprising the MAP1 family of microtubule regulatory proteins (3). However, LC3 has gained attention for MAP1-independent functions in autophagy. LC3 utilizes a ubiquitin-like conjugation system that includes E1-, E2-, and E3-like enzymes to covalently attach phosphatidylethanolamine (PE) to its C-terminus, incorporating it into the phagophore membrane during the early stages of autophagasome formation (4). Recruitment of LC3 to the phagophore may promote membrane elongation (4,5). It may also be involved in cargo recruitment to autophagosomes (1). LC3 is often used as a marker of autophagy.
- Shpilka, T. et al. (2011) Genome Biol. 12:226.
- He, H. et al. (2003) J. Biol. Chem. 278:29278.
- Kuznetsov, S.A. & V.I. Gelfand (1987) FEBS Let. 212:145.
- Weidberg, H. et al. (2011) Ann Rev. Biochem. 80:125.
- Weidberg, H. et al. (2010) EMBO J. 29:1792.
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