Human/Mouse/Rat gp96/HSP90B1 Antibody Summary
Accession # P14625
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.
Detection of Human, Mouse, and Rat gp96/HSP90B1 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, A20 mouse B cell lymphoma cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human/Mouse/Rat gp96/HSP90B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7606) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for gp96/HSP90B1 at approximately 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
gp96/HSP90B1 in Human Mesothelioma Tissue. gp96/HSP90B1 was detected in immersion fixed paraffin-embedded sections of human mesothelioma tissue using Sheep Anti-Human/Mouse/Rat gp96/HSP90B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7606) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and plasma membrane. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Western Blot Shows Human gp96/HSP90B1 Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and gp96/HSP90B1 knockout HEK293T cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human/Mouse/Rat gp96/HSP90B1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7606) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for gp96/HSP90B1 at approximately 100 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
- 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.
Glycoprotein 96 (gp96; also endoplasmin, GRP-94, TRA1 and HSP90B1) is a 94-100 kDa member of the HSP 90 family of proteins. Gp96 is a ubiquitously-expressed, ER resident protein, and is found in a preformed complex with BiP, CaBP1 and UDP-glucosyltransferase. This is a chaperone complex that binds unfolded protein substrates. When folded properly, the substrate is forwarded to calnexin-containing chaperone complexes that promote its maturation. Gp96 clients are restricted, and include disulfide-bonded integrins, TLRs, LDLR and CD180. Within the complex, gp96 exists as a disulfide-linked homodimer that may form higher-order oligomers. Gp96 also appears on the cell surface, and may serve as a receptor for bacteria. Mature human gp96 is a 782 amino acid (aa) membrane-associated protein (aa 22‑803). It is not a transmembrane protein, but utilizes an ER retention signal (aa 800-803) to interact with the ER membrane. The molecule possesses a HATPase‑C like region (aa 98-219), plus multiple ATP binding and two utilized phosphorylation sites. Over aa 503‑660, human gp96 shares 98% aa sequence identity with mouse gp96.
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