Human gp96/HSP90B1 Antibody (816803)

R&D Systems | Catalog # MAB7606

R&D Systems

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Hamster, Virus

Applications

Validated:

Knockout Validated, Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry-Frozen, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 816803
View all gp96/HSP90B1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human gp96/HSP90B1
Arg503-Arg660
Accession # P14625

Specificity

Detects human gp96/HSP90B1 in direct ELISAs and Western blots. Detects mouse and rat gp96/HSP90B1 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human gp96/HSP90B1 Antibody (816803)

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

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

Western blot shows lysates of 293T human embryonic kidney cell line, HeLa human cervical epithelial carcinoma cell line, A20 mouse B cell lymphoma cell line, CH-1 mouse B cell lymphoma cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human gp96/HSP90B1 Monoclonal Antibody (Catalog # MAB7606) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

gp96/HSP90B1 in HeLa Human Cell Line.

gp96/HSP90B1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human gp96/HSP90B1 Monoclonal Antibody (Catalog # MAB7606) at 10 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 Cells on Coverslips.
gp96/HSP90B1 antibody in Human Mesothelioma by Immunohistochemistry (IHC-P).

gp96/HSP90B1 in Human Mesothelioma.

gp96/HSP90B1 was detected in immersion fixed paraffin-embedded sections of human mesothelioma using Mouse Anti-Human gp96/HSP90B1 Monoclonal Antibody (Catalog # MAB7606) 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 plasma membranes and cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human gp96/HSP90B1 antibody by Simple WesternTM.

Detection of Human gp96/HSP90B1 by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, loaded at 0.5 mg/mL. A specific band was detected for gp96/HSP90B1 at approximately 108 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human gp96/HSP90B1 Monoclonal Antibody (Catalog # MAB7606). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western Blot Shows Human gp96/HSP90B1 Antibody Specificity by Using Knockout Cell Line.

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 0.2 µg/mL of Mouse Anti-Human gp96/HSP90B1 Monoclonal Antibody (Catalog # MAB7606) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Applications for Human gp96/HSP90B1 Antibody (816803)

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human mesothelioma

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.

Simple Western

2 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Western Blot

0.2 µg/mL
Sample: 293T human embryonic kidney cell line, HeLa human cervical epithelial carcinoma cell line, A20 mouse B cell lymphoma cell line, CH‑1 mouse B cell lymphoma cell line, and L6 rat myoblast cell line

Reviewed Applications

Read 2 reviews rated 5 using MAB7606 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 that 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

Alternate Names

ECGP, Endoplasmin, Grp94, HSP90B1

Entrez Gene IDs

7184 (Human); 22027 (Mouse); 362862 (Rat)

Gene Symbol

HSP90B1

UniProt

P14625 (Human)

Additional gp96/HSP90B1 Products

Product Documents for Human gp96/HSP90B1 Antibody (816803)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human gp96/HSP90B1 Antibody (816803)

For research use only

Citations for Human gp96/HSP90B1 Antibody (816803)

Customer Reviews for Human gp96/HSP90B1 Antibody (816803) (2)

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  • Human gp96/HSP90B1 Antibody
    Name: Armen Petrosyan
    Application: Western Blot
    Sample Tested: RM1 murine prostate cancer cells
    Species: Mouse
    Verified Customer | Posted 06/27/2023
    W-B of RM1 cell lysate
    Human gp96/HSP90B1 Antibody (816803) MAB7606
  • Human gp96/HSP90B1 Antibody
    Name: Armen Petrosyan
    Application: Western Blot
    Sample Tested: PC-3 human prostate cancer cell line
    Species: Human
    Verified Customer | Posted 06/27/2023
    W-B of PC-3 cell lysate
    Human gp96/HSP90B1 Antibody (816803) MAB7606

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Protocols

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