GRP94 Recombinant Rabbit Monoclonal Antibody [PSH25-59]
Catalog# HA724533
GRP94 Recombinant Rabbit Monoclonal Antibody [PSH25-59]
-
WB
-
IF-Cell
-
IHC-P
-
FC(Intra)
-
IP
-
Human
-
Mouse
-
Rat
-
Green monkey
-
Pig
-
HA751978
不含抗保成分
-
unconjugated
Overview
Product Name
GRP94 Recombinant Rabbit Monoclonal Antibody [PSH25-59]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Species Reactivity
Human, Mouse, Rat, Green monkey, Pig
Validated Applications
WB, IF-Cell, IHC-P, FC(Intra), IP
Target Molecular Weight
Predicted band size: 92 kDa
Positive Control
HeLa (Human cervical adenocarcinoma cell) cell lysate, MCF7 (Human breast cancer cell) cell lysate, PANC-1 (Human epidermoid carcinoma pancreatic ductal cell) cell lysate, MEF (Mouse embryonic fibroblast) cell lysate, PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate, COS-1 (African green monkey kidney fibroblast) cell lysate, Pig brain tissue lysate, Mouse brain tissue lysate, Mouse stomach tissue lysate, Rat brain tissue lysate, Rat stomach tissue lysate.
Conjugation
unconjugated
Clone Number
PSH25-59
Product Features
Form
Liquid
Concentration
0.5 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:5,000-1:10,000
-
IF-Cell
-
1:500-1:2,500
-
IHC-P
-
1:5,000
-
FC(Intra)
-
1 μg/mL
-
IP
-
1-2μg/sample
Target
Function
Heat shock protein 90kDa beta member 1 (HSP90B1), known also as endoplasmin, gp96, grp94, or ERp99, is a chaperone protein that in humans is encoded by the HSP90B1 gene. HSP90B1 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. Tumor-derived HSP90B1 (vitespen) has entered clinical trials for cancer immunotherapy. 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.
Background References
1. Xu H et al. Conformationally Restricted Grp94-Selective Inhibitors. ACS Omega. 2025 Jan
2. Ding S et al. AI662270/GRP94 axis couples the unfolded protein response to mitochondrial dynamics during acute myocardial infarction. JCI Insight. 2025 Oct
Subcellular Location
Endoplasmic reticulum lumen, Sarcoplasmic reticulum lumen, Melanosome.
Synonyms
94 kDa glucose regulated protein antibody
94 kDa glucose-regulated protein antibody
ECGP antibody
Endoplasmin antibody
Endothelial cell (HBMEC) glycoprotein antibody
ENPL_HUMAN antibody
Glucose regulated protein 94kDa antibody
gp96 antibody
gp96 homolog antibody
GRP 94 antibody
ExpandImages
-
Western blot analysis of GRP94 on different lysates with Rabbit anti-GRP94 antibody (HA724533) at 1/10,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: MCF7 (Human breast cancer cell) cell lysate
Lane 3: PANC-1 (Human epidermoid carcinoma pancreatic ductal cell) cell lysate
Lane 4: MEF (Mouse embryonic fibroblast) cell lysate
Lane 5: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate
Lane 6: COS-1 (African green monkey kidney fibroblast) cell lysate
Lane 7: Pig brain tissue lysate
Lysates/proteins at 15 µg/Lane.
Exposure time: 12 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724533, 1/10,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 92 kDa
Observed band size: 94 kDa -
Western blot analysis of GRP94 on different lysates with Rabbit anti-GRP94 antibody (HA724533) at 1/5,000 dilution.
Lane 1: Mouse brain tissue lysate
Lane 2: Mouse stomach tissue lysate
Lane 3: Rat brain tissue lysate
Lane 4: Rat stomach tissue lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 8 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724533, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 92 kDa
Observed band size: 94 kDa -
Application: Immunocytochemistry (IF-cell)
Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724533, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue). -
Application: Immunocytochemistry (IF-cell)
Species: Mouse
Sample: NIH/3T3 (Mouse fibroblast)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724533, 1/2,500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue). -
Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Kidney
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724533, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Colon
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724533, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Kidney
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724533, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Kidney
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724533, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Colon
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724533, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Colon
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724533, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Flow Cytometry (Intra)
Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724533(1 μg/mL, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature. -
Immunoprecipitation (IP)
GRP94 was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA724533 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724533 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA724533 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724533 in HeLa cell lysate
Exposure time: 7 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA724533, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 92 kDa
Observed band size: 94 kDa
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"