Human/Mouse/Rat LC3B Antibody

Recombinant Monoclonal Antibody.
  
  • 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 # Q9H492
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied 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.
Data Examples
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 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 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 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.
Knockout Validated
Western Blot Show Human LC3B SpecificityUsing Knockout Cell Line. Westernblot shows lysates of HeLa human cervical epithelial carcinoma parental cellline and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with50 μM Chloroquine for 18 hours. PVDF membrane was probed with 0.1 µg/mLof 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 (asindicated) in the parental HeLa cell line, but is not detectable in theknockout HeLa cell line. GAPDH (Catalog # AF5718)is shown as a loading control. Thisexperiment was conducted under reducing conditions and using ImmunoblotBuffer Group 1.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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:
    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
  • 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; microtubule associated protein 3 b; microtubule associated protein 3 beta; microtubule-associated protein 1 light chain 3 betaMAP1A/MAP1B light chain 3 B; microtubule-associated proteins 1A/1B light chain 3; microtubule-associated proteins 1A/1B light chain 3B
Isotype Controls
Description Application Cat# Citations Images  

Normal Rabbit IgG Control

Ctrl AB-105-C 42  
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Normal Rabbit IgG Control

Ctrl, CyTOF-ready MAB1050  
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Secondary Antibodies
Description Application Cat# Citations Images  

Rabbit IgG HRP-conjugated Antibody

WB HAF008 23  
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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL004 10
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Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL005 2
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Rabbit IgG APC-conjugated Antibody

Flow F0111 1  
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Rabbit IgG PE-conjugated Antibody

Flow F0110 2  
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Rabbit IgG Fluorescein-conjugated Antibody

Flow F0112 5  
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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL006 4
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Goat Anti-Rabbit IgG Biotinylated Antibody

WB BAF008 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC003  
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Rabbit IgG Antibody

WB AF008 1
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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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