GRP94 Rabbit Polyclonal Antibody
cat.: ER1511-5
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 92 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human GRP94 aa 194-247.
Positive control: HepG2 cell lysate, HeLa cell lysate, human brain tissue lysate, NIH/3T3 cell lysate, mouse placenta tissue lysate, mouse brain tissue lysate, PC-12 cell lysate, rat placenta tissue lysate, rat brain tissue lysate, HeLa, NIH/3T3, PC-12 , human liver tissue, human breast tissue, human stomach cancer tissue, mouse stomach tissue.
Subcellular location: Endoplasmic reticulum lumen, Melanosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:500-1:2,000
1:500
1:100-1:1,000
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
ER1511-5_1.jpg Fig1: Western blot analysis of GRP94 on different lysates with Rabbit anti-GRP94 antibody (ER1511-5) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate (10 µg/Lane)
Lane 2: HeLa cell lysate (10 µg/Lane)
Lane 3: Human brain tissue lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (10 µg/Lane)
Lane 5: Mouse placenta tissue lysate (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (20 µg/Lane)
Lane 7: PC-12 cell lysate (10 µg/Lane)
Lane 8: Rat placenta tissue lysate (20 µg/Lane)
Lane 9: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 92 kDa
Observed band size: 94 kDa

Exposure time: 25 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER1511-5) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ER1511-5_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling GRP94 with Rabbit anti-GRP94 antibody (ER1511-5) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GRP94 antibody (ER1511-5) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ER1511-5_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling GRP94 with Rabbit anti-GRP94 antibody (ER1511-5) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GRP94 antibody (ER1511-5) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ER1511-5_4.jpg Fig4: Immunocytochemistry analysis of PC-12 cells labeling GRP94 with Rabbit anti-GRP94 antibody (ER1511-5) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GRP94 antibody (ER1511-5) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ER1511-5_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-GRP94 antibody (ER1511-5) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1511-5) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1511-5_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-GRP94 antibody. Counter stained with hematoxylin.
ER1511-5_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue using anti-GRP94 antibody. Counter stained with hematoxylin.
ER1511-5_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse stomach tissue using anti-GRP94 antibody. Counter stained with hematoxylin.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.