hh Human LC3A Antibody MAB8558: R&D Systems

Human LC3A Antibody

(1 Review)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human LC3A in direct ELISAs.
  • Source
    Monoclonal Rat IgG2B Clone # 877005
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human LC3A
    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
    2 µg/mL
    See below
  • Immunohistochemistry
    8-25 µg/mL
    See below
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, Mouse, and Rat LC3A by Western Blot. Western blot shows lysates of human brain tissue, 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 2 µg/mL of Rat Anti-Human LC3A Antibody (Catalog # MAB8558) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). Specific bands were detected for LC3A at approximately 14 and 16 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry
LC3A in Human Brain Cortex Tissue. LC3A was detected in immersion fixed paraffin-embedded sections of human brain cortex tissue using Rat Anti-Human LC3A Monoclonal Antibody (Catalog # MAB8558) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS017) and counterstained with hematoxylin (blue). Specific staining was localized to neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Immunohistochemistry
LC3A in Human Brain. LC3A was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Rat Anti-Human LC3A Monoclonal Antibody (Catalog # MAB8558) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS017) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
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: LC3A

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 alpha
  • Entrez Gene IDs:
    84557 (Human); 66734 (Mouse); 362245 (Rat); 852200 (Yeast)
  • Alternate Names:
    Apg8; APG8a; Apg8p3; ATG8E; Autophagy-related protein LC3 A; Autophagy-related ubiquitin-like modifier LC3 A; LC3; LC3A; lc3-i, ii; MAP1A/1B light chain 3 A; MAP1ALC3; MAP1ALC3MAP1A/MAP1B LC3 A; MAP1BLC3MAP1A/MAP1B light chain 3 A; MAP1LC3A; microtubule-associated protein 1 light chain 3 alphaMAP1 light chain 3-like protein 1; microtubule-associated proteins 1A/1B light chain 3; microtubule-associated proteins 1A/1B light chain 3A; MLP3A
Related Research Areas
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Immunohistochemistry Reagents
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VisUCyte Antigen Retrieval Reagent-Basic

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Isotype Controls
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Rat IgG2B Isotype Control

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Normal Rat IgG Control

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Normal Rat IgG Control (Azide Free)

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Rat IgG2B Isotype Control

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Secondary Antibodies
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Rat IgG HRP-conjugated Antibody

WB, Simple Western HAF005 7  
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Goat Anti-Rat F(ab)2 IgG (H+L)
APC-conjugated Antibody

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Rat F(ab)2 IgG (H+L) PE-conjugated Antibody

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Goat Anti-Rat IgG NorthernLights™ NL493-conjugated Antibody

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Goat Anti-Rat IgG Biotinylated Antibody

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Rat IgG VisUCyte HRP Polymer Antibody

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Goat Anti-Rat IgG Antibody

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Rat F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

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Goat Anti-Rat IgG NorthernLights™ NL637-conjugated Antibody

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Goat Anti-Rat F(ab)2 IgG (H+L) PerCP-conjugated Antibody

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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Immunohistochemistry.

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 IHC Human Anonymous 08/30/2017
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Summary

ApplicationImmunohistochemistry
Sample TestedCancer cell lysates
SpeciesHuman

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