Human BAK Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects endogenous human BAK in Western blots.
  • Source
    Monoclonal Mouse IgG1 Clone # 564305
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human BAK
    Pro20-Asn124
    Accession # Q16611
  • 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
    1 µg/mL
    See below
  • Immunocytochemistry
    5-25 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 BAK by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, 293T human embryonic kidney epithelial cell line, and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human BAK Monoclonal Antibody (Catalog # MAB8161) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for BAK at approximately 23 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Immunocytochemistry
BAK in Jurkat Human Cell Line. BAK was detected in immersion fixed Jurkat human acute T cell leukemia cell line using Mouse Anti-Human BAK Monoclonal Antibody (Catalog # MAB8161) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
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: BAK
BAK (Bcl-2 homologous antagonist/killer; also know as BAK1 and Bcl-2-like protein 7) is a 25‑30 kDa member of the BCL-2 family of proteins. It is widely expressed and participates in the apoptotic cycle. BAK is an outer mitochondrial membrane protein that is inactive as a Zn-dependent homodimer. Upon activation by p53 or tBID, BAK oligomerizes, creating a pore in the mitochondrial membrane and allowing for cytochrome C release. Human BAK contains three Bcl-2 homology domains (aa 74‑88, 117‑136 and 169‑184), a Zn-binding region (aa 160‑166) and a C-terminal transmembrane segment (aa 188‑205). Amino acids 67‑94 mediate oligomerization of BAK. There are two potential isoform variants; one shows an alternate start site at Met 96, while a second shows a deletion of aa 46‑66. Over amino acids 20‑124, human BAK shares 76% aa identity with mouse BAK.
  • Long Name:
    Bcl-2 Antagonist/Killer
  • Entrez Gene IDs:
    578 (Human); 12018 (Mouse)
  • Alternate Names:
    Apoptosis regulator BAK; BAK; BAK1; BAK-LIKE; BAKMGC3887; BCL2-antagonist/killer 1; BCL2L7; bcl2-L-7; BCL2L7bcl-2 homologous antagonist/killer; BCL2-like 7 protein; Bcl-2-like protein 7; CDN1; MGC117255; pro-apoptotic protein BAK
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