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.
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.
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.
IP result of anti-GRP94 (IP:83147-5-RR, 4ug; Detection:83147-5-RR 1:5000) with HeLa cells lysate 1260 ug.
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).
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).
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).
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).
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 AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1).
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 AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1).
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 AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
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 AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
IF Staining of HEK-293 using 83147-5-RR
Immunofluorescent analysis of (-20°C Ethanol) fixed HEK-293 cells using GRP94 antibody (83147-5-RR, Clone: 230502B7 ) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).
Immunofluorescent analysis of (-20°C Ethanol) fixed HEK-293 cells using GRP94 antibody (83147-5-RR, Clone: 230502B7 ) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).
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 AffiniPure 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).
1x10^6 A431 cells were intracellularly stained with 0.25 ug Grp94 Recombinant Antibody (83147-5-RR, Clone:230502B7) and APC-Conjugated AffiniPure 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).
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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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, HEK-293 cells
Positive FC (Intra) detected in
A431 cells
Recommended dilution
Application
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.
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 -20oC 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).
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.
IHC Figures
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).
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).
IP Figures
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.
IF/ICC Figures
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 AffiniPure Goat Anti-Mouse IgG(H+L) (SA00013-1).
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 AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
IF Staining of HEK-293 using 83147-5-RR
Immunofluorescent analysis of (-20°C Ethanol) fixed HEK-293 cells using GRP94 antibody (83147-5-RR, Clone: 230502B7 ) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).
FC (INTRA) Figures
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 AffiniPure 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).
AFFINITY Figures
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.
The species listed in Tested Reactivity are in-house verified and applicable species. For unlisted species, please refer to the homology analysis of the immunogen sequence and related species. For rabbit polyclonal antibodies, homology >70% is recommended. For mouse monoclonal antibodies and rabbit recombinant antibodies, homology >90% is recommended. Generally, the higher the homology, the greater the applicability. However, there will be certain differences in protein expression in different species, tissues or cells. Therefore, the homology analysis results are for reference only and do not serve as a guarantee.
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Proteintech
GRP94 Recombinant antibody
Catalog Number
83147-5-RR
Citations
-
Dilutions
WB : 1:5000-1:50000 IP : 0.5-4.0 ug for IP and 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate for WB IHC : 1:200-1:800 IF/ICC : 1:125-1:500 FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension
Applications
WB, IHC, IF/ICC, FC (Intra), IP, ELISA
Reactivity
human, pig
Product Guarantee
Covers any species including not listed on datasheet
Covers any applications including not listed on datasheet