GRP94 Recombinant Rabbit Monoclonal Antibody [PSH25-59]
cat.: HA724533
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat, Green monkey, Pig |
| Applications: | WB, IF-Cell, IHC-P, FC(Intra), IP |
| Clonality: | Monoclonal |
| Clone number: | PSH25-59 |
| Form: | Liquid |
| Storage condition: | 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. |
| Concentration: | 0.5ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 92 kDa |
| Isotype: | IgG |
| 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. |
| Subcellular location: | Endoplasmic reticulum lumen, Sarcoplasmic reticulum lumen, Melanosome. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC(Intra) IP |
1:5,000-1:10,000 1:500-1:2,500 1:5,000 1 μg/mL 1-2μg/sample |
| Uniprot #: | SwissProt: P14625 Human | P08113 Mouse | Q66HD0 Rat |
| Alternative names: | 94 kDa glucose regulated protein 94 kDa glucose-regulated protein ECGP Endoplasmin Endothelial cell (HBMEC) glycoprotein ENPL_HUMAN Glucose regulated protein 94kDa gp96 gp96 homolog GRP 94 GRP-94 Heat shock protein 90 kDa beta member 1 heat shock protein 90kDa beta (Grp94), member 1 Heat shock protein, 90 kDa, beta, 1 HSP90B1 Stress inducible tumor rejection antigen GP96 TRA1 tumor rejection antigen (gp96) 1 Tumor rejection antigen 1 Tumor rejection antigen gp96 Tumor rejection antigen-1 (gp96) |
Images
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Fig1:
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 |
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Fig2:
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 |
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Fig3:
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). |
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Fig4:
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). |
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Fig5:
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. |
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Fig6:
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. |
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Fig7:
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. |
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Fig8:
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. |
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Fig9:
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. |
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Fig10:
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. |
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Fig11:
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. |
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Fig12:
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 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.