Human Bad Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Bad in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant mouse (rm) Bad (aa 1-130), rmBcl-2 alpha, recombinant human (rh) Bcl-2, or rhBCL2L12 is observed.
  • Source
    Monoclonal Mouse IgG2A Clone # 612312
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human Bad
    Met1-Gln168
    Accession # Q92934
  • 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
    0.5 µg/mL
    See below
  • Immunohistochemistry
    8-25 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, Mouse, and Rat Bad by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line, Neuro‑2A mouse neuroblastoma cell line, and NRK rat normal kidney cell line. PVDF Membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat Bad Monoclonal Antibody (Catalog # MAB6405) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Bad at approximately 23 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Immunocytochemistry
Bad in Jurkat Human Cell Line. Bad was detected in immersion fixed Jurkat human acute T cell leukemia cell line using Mouse Anti-Human Bad Monoclonal Antibody (Catalog # MAB6405) at 25 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 Non-adherent Cells.
Immunohistochemistry
Bad in Human Kidney. Bad was detected in immersion fixed paraffin-embedded sections of human kidney using Mouse Anti-Human/Mouse/Rat Bad Monoclonal Antibody (Catalog # MAB6405) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of epithelial cells in convoluted tubules. 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: Bad
Bcl-2 antagonist of cell death (Bad) is a 23 kDa cytoplasmic protein in the Bcl-2 family. It functions as a pro-apoptotic molecule by dimerizing with and inhibiting the anti-apoptotic proteins Bcl-2 and Bcl-xL. Prosurvival signals trigger the phosphorylation of Bad on Ser115, disrupting its interaction with Bcl-2 and Bcl-xL and resulting in protection from apoptosis. Human Bad shares 75% aa sequence identity with mouse and rat Bad.
  • Long Name:
    Bcl-xL/Bcl-2 Associated Death Promoter
  • Entrez Gene IDs:
    572 (Human); 12015 (Mouse); 64639 (Rat)
  • Alternate Names:
    Bad; BBC2; BBC6; bcl2 antagonist of cell death; BCL2-antagonist of cell death protein; BCL2-associated agonist of cell death; Bcl-2-binding component 6; BCL2-binding component 6; BCL2-binding protein; BCL2L8; Bcl2-L-8; BCL2L8bcl2-L-8; Bcl-2-like protein 8; BCL-X/BCL-2 binding protein; Bcl-XL/Bcl-2-associated death promoter
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