Human/Mouse/Rat LC3B Antibody

Catalog #: MAB85581 Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB85581-100
MAB85581-SP

Detection of Human LC3B by Western Blot.

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Human/Mouse/Rat LC3B Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human, mouse, and rat LC3B in Western blots.

Source

Recombinant Monoclonal Rabbit IgG Clone # 1251D

Purification

Protein A or G purified from cell culture supernatant

Immunogen

Human LC3B synthetic peptide
Accession # Q9GZQ8

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

See below

Simple Western

5 µg/mL

See below

Knockout Validated

LC3B is specifically detected in HeLa human cervical epithelialcarcinoma parental cell line treated with Chloroquine but is not detectable inLC3B knockout HeLa cell line treated with Chloroquine.

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

Western Blot

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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.

Western Blot

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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.

Simple Western

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Detection of Mouse LC3B by Simple Western^TM. 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.

Simple Western

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Detection of Rat LC3B by Simple Western^TM. 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.

Knockout Validated

Western Blot Show Human LC3B Antibody Specificity Using Knockout Cell Line.

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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

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

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.

References

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.

Long Name

Microtubule-associated Protein 1 Light Chain 3 beta

Entrez Gene IDs

81631 (Human); 67443 (Mouse); 64862 (Rat)

Alternate Names

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