gp96/HSP90B1/GRP94 Antibody [Unconjugated]

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 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 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.

• Reactivity: Hu, Mu, Rt
• Applications: WB, IHC, KO
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

gp96/HSP90B1/GRP94 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human gp96/HSP90B1
  Arg503-Arg660
  Accession # P14625
• Specificity: Detects human, mouse, and rat gp96/HSP90B1 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: HSP90B1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 1-15 ug/mL
  • Knockout Validated
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for gp96/HSP90B1/GRP94 Antibody [Unconjugated]

• 94 kDa glucose-regulated protein
• ECGP
• Endoplasmin
• endothelial cell (HBMEC) glycoprotein
• glucose regulated protein, 94 kDa
• gp96
• GP96ECGP
• Grp94
• GRP-94
• GRP94gp96 homolog
• Heat shock protein 90 kDa beta member 1
• heat shock protein 90kDa beta (Grp94), member 1
• HSP90B1
• TRA1endoplasmin
• tumor rejection antigen (gp96) 1
• Tumor rejection antigen 1
• Tumor rejection antigen-1 (gp96)

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for gp96/HSP90B1/GRP94 Antibody (AF7606)(2)

Publications using AF7606

• Laura Padula, Eva Fisher, Katelyn Rivas, Kristin Podack, Daniela Frasca, Jonah Kupritz et al. Secreted heat shock protein gp96-Ig and OX40L-Fc combination vaccine enhances SARS-CoV-2 Spike (S) protein-specific B and T cell immune responses Vaccine: X 2022-12-01 [PMID: 35936992] (ELISA Capture, Human)
• KA Piróg, EP Dennis, CL Hartley, RM Jackson, J Soul, JM Schwartz, JF Bateman, RP Boot-Handf, MD Briggs XBP1 signalling is essential for alleviating mutant protein aggregation in ER-stress related skeletal disease PLoS Genet., 2019-07-01;15(7):e1008215. 2019-07-01 [PMID: 31260448] (IHC, Mouse)

Bioinformatics

• Gene Symbol: HSP90B1
• Uniprot:
  • P14625(Human)