WB result of GRP94 Rabbit mAb
Primary antibody: GRP94 Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: MCF7 whole cell lysate 20 µg
Lane 3: PANC-1 whole cell lysate 20 µg
Lane 4: MOLT-4 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 92 kDa
Observed MW: 100 kDa
Product Details
Product Details
Product Specification
Host | Rabbit |
Antigen | GRP94 |
Synonyms | Endoplasmin, 94 kDa glucose-regulated protein (GRP-94), Heat shock protein 90 kDa beta member 1, Tumor rejection antigen 1, gp96 homolog, HSP90B1, TRA1 |
Location | Melanosome |
Accession | P14625 |
Clone Number | S-R268 |
Antibody Type | Recombinant mAb |
Isotype | IgG |
Application | WB, IHC-P, ICC, IP, ICFCM |
Reactivity | Hu, Ms, Rt |
Purification | Protein A |
Concentration | 0.5 mg/ml |
Conjugation | Unconjugated |
Physical Appearance | Liquid |
Storage Buffer | PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300 |
Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied. |
Dilution
application | dilution | species |
WB | 1:1000-1:10000 | |
IP | 1:50 | |
IHC-P | 1:250 | |
ICC | 1:500 | |
ICFCM | 1:500 |
Background
GRP94 is an HSP90 paralogue that is found in the endoplasmic reticulum. It plays critical roles in folding proteins in the secretory pathway such as Toll-like receptors and integrins. It has been implicated as an essential immune chaperone to regulate both innate and adaptive immunity. Grp94 has been shown to be a target for treatment of a plethora of diseases such as glaucoma, multiple myeloma, and metastatic cancer. grp94 includes 5 distinct amino acids in its primary sequence which creates 2 unique sub-pockets, S1 and S2. These sub-pockets have been utilized in current research in order to inhibit the chaperone since its client proteins seem to be up-regulated in cancer cells.
Picture
Picture
Western Blot
WB result of GRP94 Rabbit mAb
Primary antibody: GRP94 Rabbit mAb at 1/1000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 92 kDa
Observed MW: 100 kDa
WB result of GRP94 Rabbit mAb
Primary antibody: GRP94 Rabbit mAb at 1/1000 dilution
Lane 1: PC-12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 92 kDa
Observed MW: 100 kDa
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling GRP94 antibody at 1/500 (0.1 μg) dilution / (red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
IP
GRP94 Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating GRP94 in 0.4 mg HeLa whole cell lysate.
Western blot was performed on the immunoprecipitate using GRP94 Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1: HeLa whole cell lysate 20 µg (Input)
Lane 2: GRP94 Rabbit mAb IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa whole cell lysate
Predicted MW: 94 kDa
Observed MW: 94 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human breast cancer. Anti-GRP94 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human endometrial cancer. Anti-GRP94 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung squamous cell carcinoma. Anti-GRP94 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human prostatic cancer. Anti-GRP94 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse colon. Anti-GRP94 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat stomach. Anti-GRP94 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti-GRP94 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).
