gp96/HSP90B1/GRP94 Antibody (9G10)

Western Blot: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Analysis of 25 ug of Hela (lane 1), C6 (lane 2) and NIH-3T3 (lane 3) lysates.

Immunocytochemistry/Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - GR94 in human primary fibroblasts, ER staining. Primary antibody incubation: 1:250 in 10% Serum in PBST (0.1% Triton) O/N 4C. Secondary: Donkey anti-rat 647 1:500 2h RT. ICC/IF image submitted by a verifed customer review.

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Normal biopsies of deparaffinized Mouse breast tissue.

Western Blot: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Analysis of 2-fold serial dilutions of HeLa cell lysate, starting at 10 ug, per well.

Immunocytochemistry/Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Analysis of Glucose-Regulated Protein 94 using Glucose-Regulated Protein 94 Monoclonal Antibody (9G10) shows staining in C6 Cells. Glucose-Regulated Protein 94 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Glucose-Regulated Protein 94 at a dilution of 1:100 over night at 4C, washed with PBS and incubated with a DyLight-488 conjugated.

Immunocytochemistry/Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Analysis of Glucose-Regulated Protein 94 using Glucose-Regulated Protein 94 Monoclonal Antibody (9G10) shows staining in Hela Cells. Glucose-Regulated Protein 94 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Glucose-Regulated Protein 94 at a dilution of 1:100 over night at 4C, washed with PBS and incubated with a DyLight-488 conjugated.

Immunocytochemistry/Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Analysis of Glucose-Regulated Protein 94 using Glucose-Regulated Protein 94 Monoclonal Antibody (9G10) shows staining in NIH-3T3 Cells. Glucose-Regulated Protein 94 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Glucose-Regulated Protein 94 at a dilution of 1:20 over night at 4C, washed with PBS and incubated with a DyLight-488 conjugated.

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Normal biopsies of deparaffinized Mouse liver tissue.

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Normal biopsies of deparaffinized Mouse lymph node.

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Analysis of Glucose-Regulated Protein 94 (Grp94) in U2OS cells. Cells fixed with 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature and blocked with 2% BSA in PBST (from Product # 37525) for 30 minutes at room temperature. Cells were treated with Peroxidase Suppressor, probed with a Grp94 monoclonal antibody at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBST, and incubated with an HRP-conjugated goat anti-rat IgG (H+L) secondary antibody at a dilution of 1:1000 for 30 minutes at room temperature. Chromogenic detection was performed using Metal Enhanced DAB Substrate Kit.

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] - Staining on poorly differentiated adenocarcinoma from paraffin-embedded human colon tissue sections.

• Reactivity: Hu, Mu, Rt, Bv, Ch, Ha
• Applications: WB, ICC/IF, IHC, IHC-P, IP, ICC/IF
• Clone: 9G10
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated

gp96/HSP90B1/GRP94 Antibody (9G10) Summary

• Immunogen: Chick oviduct GRP94.
• Localization: Endoplasmic reticulum
• Marker: ER Stress Marker
• Specificity: GRP94 (9G10)
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Gene: HSP90B1
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry
  • Immunocytochemistry/Immunofluorescence 1:250
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:20
  • Immunoprecipitation 1:10 - 1:500
  • Western Blot 1:100 - 1:1000
• Application Notes: WB: Detects an approx. 108 kDa protein representing GRP94 in chick oviduct cytosol.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: Ascites
• Preservative: 0.05% Sodium Azide
• Purity: Unpurified

Alternate Names for gp96/HSP90B1/GRP94 Antibody (9G10)

• 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

Glucose-regulated protein 94, also known as Grp94 or gp96, is an abundant resident endoplasmic reticulum (ER) lumenal stress protein which together with cytosolic Hsp90 belongs to the Hsp90 family of molecular chaperones. Grp94 and other resident soluble proteins of the ER such as members of the Ca(2+) binding protein subfamily (CaBP), CaBPI and CaBP2 as well as calreticulin, possess the COOH-terminal tetrapeptide Lys-Asp-Glu-Leu (KDEL) which is a sorting signal that is thought to lead to the retention of these proteins in the pre-Golgi compartments (1). Grp94 expression is upregulated by stress conditions such as lucose starvation and heat shock, which promote protein misfolding or unfolding (2). In addition to a homeostatic role in protein folding and assembly, Grp94 can function in the intracellular trafficking of peptides from the extracellular space to the MHC class I antigen processing pathway of antigen presentation cells (3,4). Grp94 and Hsp90 share high sequence identity and presumably identical adenosine nucleotide-dependent modes of regulation. Earlier data suggests that Hsp90 and Grp94 may differ in their nucleotide binding properties. The N-terminal domain of eukaryotic Hsp90 proteins contains a conserved adenosine nucleotide binding pocket which also serves as the inding site for the Hsp90 inhibitors geldanamycin and radicicol. However, the molecular basis for adenosine nucleotide-dependent regulation of Grp94remains to be established. Recent data has entified a ligand dependent regulation of Grp94 function and suggest a model whereby Grp94 function is regulated through a ligand-dependent conversion of Grp94 from an inactive to an active conformation (5, 6).

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 (NB300-619)(1)

Publication using NB300-619

• Vierra NC, Dickerson MT, Jordan KL et al. TALK-1 reduces delta-cell endoplasmic reticulum and cytoplasmic calcium levels limiting somatostatin secretion. Mol Metab 2018 Mar [PMID: 29402588]

Bioinformatics

• Gene Symbol: HSP90B1
• Entrez:
  • 374163(Chicken)
  • 22027(Mouse)
• Uniprot:
  • P08110(Chicken)