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Human/Mouse/Rat BAK Antibody

R&D Systems, part of Bio-Techne | Catalog # AF8161

R&D Systems, part of Bio-Techne
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AF8161
AF8161-SP

Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Immunocytochemistry, Immunohistochemistry, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

E. coli-derived recombinant human BAK
Pro20-Asn124
Accession # Q16611

Specificity

Detects endogenous human, mouse and rat BAK in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat BAK Antibody

Detection of Human/Mouse/Rat BAK antibody by Western Blot.

Detection of Human/Mouse/Rat BAK by Western Blot.

Western blot shows lysates of HEK293 human embryonic kidney cell line, CH-1 mouse B cell lymphoma cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat BAK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8161) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for BAK at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
BAK antibody in HEK293 Human Cell Line by Immunocytochemistry (ICC).

BAK in HEK293 Human Cell Line.

BAK was detected in immersion fixed HEK293 human embryonic kidney cell line using Goat Anti-Human/Mouse/Rat BAK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8161) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of BAK in Human Colon.

BAK was detected in immersion fixed paraffin-embedded sections of Human Colon using Goat Anti-Human/Mouse/Rat BAK Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8161) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membrane in epithelial cells in mucosal glands. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Applications for Human/Mouse/Rat BAK Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HEK293 human embryonic kidney cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of Human Colon

Simple Western

4 µg/mL
Sample: Jurkat human acute T cell leukemia cell line and THP-1 human acute monocytic leukemia cell line

Western Blot

1 µg/mL
Sample: HEK293 human embryonic kidney cell line, CH-1 mouse B cell lymphoma cell line, and PC-12 rat adrenal pheochromocytoma cell line

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*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.

Background: BAK

BAK (Bcl-2 homologous antagonist/killer; also BAK1) 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 is 211 amino acids (aa) in length and 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 Met96, 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

Alternate Names

BAK1, BCL2L7

Entrez Gene IDs

578 (Human); 12018 (Mouse)

Gene Symbol

BAK1

UniProt

Additional BAK Products

Product Documents for Human/Mouse/Rat BAK Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse/Rat BAK Antibody

For research use only

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