gp96/HSP90B1/GRP94 Antibody (9G10)

Novus Biologicals | Catalog # NB300-619

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Bovine, Chicken, Hamster

Cited:

Human

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2A Clone # 9G10
View all gp96/HSP90B1 Primary Antibodies »

Product Specifications

Immunogen

Chick oviduct GRP94.

Localization

Endoplasmic reticulum

Marker

ER Stress Marker

Specificity

GRP94 (9G10)

Clonality

Monoclonal

Host

Rat

Isotype

IgG2A

Scientific Data Images for gp96/HSP90B1/GRP94 Antibody (9G10)

Western Blot: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

Western Blot: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Immunocytochemistry/ Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Western Blot: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Immunocytochemistry/ Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Immunocytochemistry/ Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Immunocytochemistry/ Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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]

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619]

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

Western Blot: gp96/HSP90B1/GRP94 Antibody (9G10) [NB300-619] -

Isolation and characterization of EVs. (A) Schematic representation of the ultracentrifugation protocol employed to isolate distinct EVs subpopulations. The pellets collected at 2K, 10K, 100K, and 200 K were sequentially washed in PBS at the respective centrifugation speeds and resuspended to a concentration of 1 million cells per uL. Image generated using BioRender.com. (B) Diagram of the SEC method used for EVs isolation. A total of 25, 0.5 mL fractions were collected for further analysis. Image generated using BioRender.com. (C) Western blot analysis showing the expression of common EVs surface markers—CD63, CD9, and CD81—confirms EVs’ presence. The absence of gp96 in the EVs lanes indicates minimal contamination from other cellular components. The total cellular homogenate was used as a control. (D) Dot blot analysis of the 25 fractions obtained from SEC, probing for CD63 to identify fractions containing EVs. Fractions containing EVs are boxed and numbered accordingly. (E) EVs morphology analysis by TEM. EVs display a typical cup-shaped appearance, consistent with the preparation process and negative staining. The scale bar is 50 nm. (F) NTA results depict the concentration and size distribution of EVs recovered by SEC, with a particle size range consistent with extracellular vesicles. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40567500), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for gp96/HSP90B1/GRP94 Antibody (9G10)

Application
Recommended Usage

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.

Reviewed Applications

Read 2 reviews rated 4.5 using NB300-619 in the following applications:

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Ascites

Preservative

0.05% Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: gp96/HSP90B1

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).

Long Name

Heat Shock Protein 90 beta 1

Alternate Names

ECGP, Endoplasmin, Grp94, HSP90B1

Entrez Gene IDs

22027 (Mouse); 374163 (Chicken)

Gene Symbol

HSP90B1

UniProt

P08110 (Chicken)

Additional gp96/HSP90B1 Products

Product Documents for gp96/HSP90B1/GRP94 Antibody (9G10)

Certificate of Analysis

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Product Specific Notices for gp96/HSP90B1/GRP94 Antibody (9G10)

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.

Citations for gp96/HSP90B1/GRP94 Antibody (9G10)

Customer Reviews for gp96/HSP90B1/GRP94 Antibody (9G10) (2)

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Customer Images

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  • gp96/HSP90B1/GRP94 Antibody (9G10)
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Human primary fibroblast
    Species: Human
    Verified Customer | Posted 11/22/2020
    GR94 in human primary fibroblasts, ER staining. Primary ab incubation: 1:250 in 10% Serum in PBST (0.1% Triton) O/N 4ºC. Secondary: Donkey anti-rat 647 1:500 2h RT.
    PFA 4% 15 min fixed. Primary ab incubation: 1:250 in 10% Serum in PBST (0.1% Triton) O/N 4ºC. Secondary: Donkey anti-rat 647 1:500 2h RT.
    gp96/HSP90B1/GRP94 Antibody (9G10) NB300-619
  • gp96/HSP90B1/GRP94 Antibody (9G10)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Human primary fibroblast
    Species: Human
    Verified Customer | Posted 10/25/2020
    GRP94 in human primary fibroblasts samples after a protein gradient prep. Primary ab dilution 1:1000 in BSA 3% in PBST (0.1 tween), O/N incubation, 4ºC. Secondary ab: Donkey anti-rat Alexa fluor 555.
    gp96/HSP90B1/GRP94 Antibody (9G10) NB300-619

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