Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Mouse, Bovine

Applications

Validated:

Knockout Validated, Western Blot, Immunocytochemistry, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 1251B
View all LC3B Primary Antibodies »

Product Specifications

Immunogen

Human LC3B synthetic peptide
Accession # Q9GZQ8

Specificity

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

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat LC3B Antibody

Western Blot Show Human LC3B Specificity Using Knockout Cell Line.

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 50uM Chloroquine for 18 hours. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat LC3B Monoclonal Antibody (Catalog # MAB85582) 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.
Detection of Human LC3B antibody by Western Blot.

Detection of Human LC3B by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 50uM Chloroquine for 18 Hours. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat LC3B Monoclonal Antibody (Catalog # MAB85582) 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 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 50uM Chloroquine for 18 hours. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat LC3B Monoclonal Antibody (Catalog # MAB85582) 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.
LC3B antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

LC3B in HeLa Human Cell Line.

LC3B was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with Chloroquine using Rabbit Anti-Human/Mouse/Rat LC3B Monoclonal Antibody (Catalog # MAB85582) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Mouse LC3B antibody by Simple WesternTM.

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 50uM 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/Mouse/Rat LC3B Monoclonal Antibody (Catalog # MAB85582). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for Human/Mouse/Rat LC3B Antibody

Application
Recommended Usage

Immunocytochemistry

1-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line treated with Chloroquine

Knockout Validated

0.1 µg/mL
Sample: HeLa human cervical epithelial carcinoma parental cell line and LC3B knockout HeLa cell line treated with Chloroquine.

Simple Western

5 µg/mL
Sample: NIH‑3T3 mouse embryonic fibroblast cell line treaed with Chloroquine

Western Blot

0.1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and PC‑12 rat adrenal pheochromocytoma cell line treated with Chloroquine

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

  1. Shpilka, T. et al. (2011) Genome Biol. 12:226.
  2. He, H. et al. (2003) J. Biol. Chem. 278:29278.
  3. Kuznetsov, S.A. & V.I. Gelfand (1987) FEBS Let. 212:145.
  4. Weidberg, H. et al. (2011) Ann Rev. Biochem. 80:125.
  5. Weidberg, H. et al. (2010) EMBO J. 29:1792. 
 

Long Name

Microtubule-associated Protein 1 Light Chain 3 beta

Alternate Names

Apg8b, ATG8F, LC3II, MAP1LC3B

Entrez Gene IDs

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

Gene Symbol

MAP1LC3B

UniProt

Q9GZQ8

Additional LC3B Products

Product Documents for Human/Mouse/Rat LC3B Antibody

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Product Specific Notices for Human/Mouse/Rat LC3B Antibody

For research use only

Citations for Human/Mouse/Rat LC3B Antibody

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Protocols

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