GRP94 Recombinant antibody

GRP94 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, pig

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

CloneNo.

230502B7

Cat no : 83147-5-RR

Synonyms

HSP90B1, Heat shock protein family C member 4, GP96, Endoplasmin, ECGP

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Validation Data Gallery

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  WB analysis using 83147-5-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83147-5-RR (GRP94 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 83147-5-RR

  IP result of anti-GRP94 (IP:83147-5-RR, 4ug; Detection:83147-5-RR 1:5000) with HeLa cells lysate 1260 ug.

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  IHC staining of human lung cancer using 83147-5-RR

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 83147-5-RR (GRP94 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 83147-5-RR

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 83147-5-RR (GRP94 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 83147-5-RR

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using GRP94 antibody (83147-5-RR, Clone: 230502B7 ) at dilution of 1:250 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  IF Staining of HeLa using 83147-5-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using GRP94 antibody (83147-5-RR, Clone: 230502B7 ) at dilution of 1:300 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1).

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  FC experiment of A431 using 83147-5-RR

  1x10^6 A431 cells were intracellularly stained with 0.25 ug Grp94 Recombinant Antibody (83147-5-RR, Clone:230502B7) and APC-Conjugated Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 83147-5-RR

  Biolayer interferometry (BLl) kinetic assays of 83147-5-RR against Human GRP94 were performed. The affinity constant is below 1 pM.

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, Jurkat cells, HepG2 cells, LNCaP cells, pig brain tissue
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human lung cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: A431 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)/ICC: IF/ICC : 1:125-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

83147-5-RR targets GRP94 in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, pig samples.

• Tested Reactivity: human, pig
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: GRP94 fusion protein Ag1439
• Full Name: heat shock protein 90kDa beta (Grp94), member 1
• Calculated Molecular Weight: 96 kDa
• Observed Molecular Weight: 100 kDa
• GenBank Accession Number: BC009195
• Gene Symbol: GRP94
• Gene ID (NCBI): 7184
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Glucose-regulated protein 94 (GRP94), also called HSP90B1 and GP96, is an endoplasmic reticulum (ER)-resident member of the heat shock protein 90 (HSP90) family (PMID: 33802964; 24658275). Under ER stress conditions, GRP94 accelerates its function as a molecular chaperone. Client proteins of GRP94 include toll-like receptors (TLRs), glycoprotein (GP) IX subunit, insulin-like growth factors (IGFs), proinsulin, and integrins (PMID: 7913987; 32781621; 22079671). As well as being an ER chaperone, GRP94 can also participate in Calcium regulation and is also a regulator of innate and adaptive immunity (PMID: 33802964; 11584270).