gp96/HSP90B1/GRP94 Antibody (CL2647) - BSA Free

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining in human prostate and skeletal muscle tissues using NBP2-42379 antibody. Corresponding HSP90B1 RNA-seq data are presented for the same tissues.

Western Blot: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Analysis in U-251MG cells transfected with control siRNA, target specific siRNA probe #1 and #2, using Anti-HSP90B1 antibody. Remaining relative intensity is presented. Loading control: Anti-PPIB.

Immunocytochemistry/Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining in U251 cell line showing specific staining of endoplasmic reticulum in green. Microtubule-staining and nuclear probes are visualized in red and blue respectively (where available). Antibody staining is shown in green. Antibody staining is shown in green.

Western Blot: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Analysis in human cell line HeLa, human cell line HEK 293, human cell line A-431 and human cell line HepG2.

Immunocytochemistry/Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining in HeLa cell line showing specific staining of endoplasmic reticulum in green. Microtubule-staining and nuclear probes are visualized in red and blue respectively (where available). Antibody staining is shown in green. Antibody staining is shown in green.

Immunocytochemistry/Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining of human cell line A431 showing specific staining of endoplasmic reticulum in green. Microtubule-staining and nuclear probes are visualized in red and blue respectively (where available). Antibody staining is shown in green. Antibody staining is shown in green.

Immunocytochemistry/Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining in MCF7 cell line showing specific staining of endoplasmic reticulum in green. Microtubule-staining and nuclear probes are visualized in red and blue respectively (where available). Antibody staining is shown in green. Antibody staining is shown in green.

Immunocytochemistry/Immunofluorescence: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining in U2OS cell line with Anti-HSP90B1 monoclonal antibody, showing specific staining of endoplasmic reticulum in green. Microtubule-staining and nuclear probes are visualized in red and blue respectively (where available). Antibody staining is shown in green. Antibody staining is shown in green.

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining of human cerebral cortex shows strong cytoplasmic positivity in neurons.

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining of human placenta shows strong cytoplasmic positivity in trophoblastic cells.

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining of human prostate shows strong cytoplasmic positivity in glandular cells.

Immunohistochemistry-Paraffin: gp96/HSP90B1/GRP94 Antibody (CL2647) [NBP2-42379] - Staining of human skeletal muscle shows very weak cytoplasmic positivity in myocytes.

gp96/HSP90B1/GRP94 (CL2647) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti-gp96/HSP90B1/GRP94 (CL2647) Protein A Purified Monoclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NBP2-42379) (light blue) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.

• Reactivity: Hu, Mu
• Applications: WB, ICC/IF, IHC, Mycoplasma
• Clone: CL2647
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free

gp96/HSP90B1/GRP94 Antibody (CL2647) - BSA Free Summary

• Description: Novus Biologicals Mouse gp96/HSP90B1/GRP94 Antibody (CL2647) - BSA Free (NBP2-42379) is a monoclonal antibody validated for use in IHC, WB and ICC/IF. Anti-gp96/HSP90B1/GRP94 Antibody: Cited in 3 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: This antibody was developed using a recombinant protein derived from P14625, with the exact immunogen sequence remaining proprietary.
• Predicted Species: Mouse (90%). Backed by our 100% Guarantee.
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: HSP90B1
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 2-10 ug/ml
  • Immunohistochemistry 1:5000 - 1:10000
  • Immunohistochemistry-Paraffin 1:5000 - 1:10000
  • Knockdown Validated
  • Western Blot 1 ug/ml
• Application Notes: For IHC-Paraffin, HIER pH 6 retrieval is recommended. ICC/IF Fixation Permeabilization: Use PFA/Triton X-100.
• Control Peptide:

  gp96/HSP90B1/GRP94 Recombinant Protein Antigen (NBP2-42379PEP)

Reactivity Notes

Expected species cross reactivity based on sequence identity: Rat (87%). Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Mouse-On-Mouse blocking reagent may be needed for IHC and ICC experiments to reduce high background signal. You can find these reagents under catalog numbers PK-2200-NB and MP-2400-NB. Please contact Technical Support if you have any questions

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH 7.2) and 40% Glycerol
• Preservative: 0.02% Sodium Azide
• Purity: Protein A purified

Alternate Names for gp96/HSP90B1/GRP94 Antibody (CL2647) - BSA Free

• 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 (NBP2-42379)(4)

Publications using NBP2-42379

• Lee J, Hwang H, Kang D et al. Therapy-induced senescent cancer cells contribute to cancer recurrence by providing a PD-L1 umbrella regulated by ribophorin 1 Research Square 2023-10-09 (ICC/IF, Human)
• Zhang X, Huang X, Xu J Et Al. NEK2 inhibition triggers anti-pancreatic cancer immunity by targeting PD-L1 Nature communications 2021-07-27 [PMID: 34315872] (WB)
• LC Chan, CW Li, W Xia, JM Hsu, HH Lee, JH Cha, HL Wang, WH Yang, EY Yen, WC Chang, Z Zha, SO Lim, YJ Lai, C Liu, J Liu, Q Dong, Y Yang, L Sun, Y Wei, L Nie, JL Hsu, H Li, Q Ye, MM Hassan, HM Amin, AO Kaseb, X Lin, SC Wang, MC Hung IL-6/JAK1 pathway drives PD-L1 Y112 phosphorylation to promote cancer immune evasion J. Clin. Invest., 2019-07-15;130(0):. 2019-07-15 [PMID: 31305264]
• Cha JH, Yang WH, Xia W et al. Metformin Promotes Antitumor Immunity via Endoplasmic-Reticulum-Associated Degradation of PD-L1 Mol. Cell 2018-08-16 [PMID: 30118680] (ICC/IF, Human)

Bioinformatics

• Gene Symbol: HSP90B1