Human/Mouse/Rat gp96/HSP90B1 Antibody

Catalog # Availability Size / Price Qty
AF7606
AF7606-SP
Detection of Human, Mouse, and Rat gp96/HSP90B1 by Western Blot.
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Product Details
Citations (2)
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Human/Mouse/Rat gp96/HSP90B1 Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse, and rat gp96/HSP90B1 in direct ELISAs and Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human gp96/HSP90B1
Arg503-Arg660
Accession # P14625
Formulation
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.5 µg/mL
See below
Immunohistochemistry
1-15 µg/mL
See below
Knockout Validated
Gp96/HSP90B1 is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in gp96/HSP90B1 knockout HEK293T cell line.
 

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.

Scientific Data

Western Blot Detection of Human, Mouse, and Rat gp96/HSP90B1 antibody by Western Blot. View Larger

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.

Immunohistochemistry gp96/HSP90B1 antibody in Human Mesothelioma Tissue by Immunohistochemistry (IHC-P). View Larger

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.

Knockout Validated Western Blot Shows Human gp96/HSP90B1 Antibody Specificity by Using Knockout Cell Line. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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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: gp96/HSP90B1

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.

Long Name
Heat Shock Protein 90 beta 1
Entrez Gene IDs
7184 (Human); 22027 (Mouse); 362862 (Rat)
Alternate Names
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)

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Citations for Human/Mouse/Rat gp96/HSP90B1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. 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
    Authors: Laura Padula, Eva Fisher, Katelyn Rivas, Kristin Podack, Daniela Frasca, Jonah Kupritz et al.
    Vaccine: X
  2. XBP1 signalling is essential for alleviating mutant protein aggregation in ER-stress related skeletal disease
    Authors: KA Piróg, EP Dennis, CL Hartley, RM Jackson, J Soul, JM Schwartz, JF Bateman, RP Boot-Handf, MD Briggs
    PLoS Genet., 2019-07-01;15(7):e1008215.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC

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