Product Type: | Rabbit polyclonal IgG, primary antibodies |
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Species reactivity: | Human, Mouse, Rat |
Applications: | WB, IF-Cell, IHC-P, FC |
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, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Concentration: | 1ug/ul |
Purification: | Immunogen affinity purified. |
Molecular weight: | Predicted band size: 74 kDa |
Isotype: | IgG |
Immunogen: | Synthetic peptide within C-terminal human Grp75. |
Positive control: | HeLa cell lysate, K-562 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse liver tissue lysate, mouse kidney tissue lysate, rat liver tissue lysate, HeLa, NIH/3T3, human liver cancer tissue, human liver tissue, mouse liver tissue, rat liver tissue, HepG2. |
Subcellular location: | Mitochondrion, nucleolus. |
Recommended Dilutions:
WB IF-Cell IHC-P FC |
1:500-1:2,000 1:200-1:500 1:50-1:200 1:50-1:100 |
Uniprot #: | SwissProt: P38646 Human | P38647 Mouse | P48721 Rat |
Alternative names: | 75 kDa glucose regulated protein 75 kDa glucose-regulated protein CSA Glucose Regulated Protein Grp 75 GRP-75 GRP75 GRP75_HUMAN Heat shock 70 kDa protein 9 Heat shock 70kD protein 9 heat shock 70kDa protein 9 Heat shock 70kDa protein 9B Heat shock protein 74 kDa A Heat shock protein A Heat shock protein cognate 74 Hsc74 Hsp74 Hsp74a HSPA9 Hspa9a HSPA9B MGC4500 mitochondrial Mortalin 2 Mortalin Mortalin perinuclear Mortalin2 MOT 2 MOT MOT2 Mthsp70 p66 mortalin P66 MOT PBP74 Peptide binding protein 74 Peptide-binding protein 74 Stress 70 protein mitochondrial Stress 70 protein mitochondrial precursor Stress-70 protein |
Fig1:
Western blot analysis of Grp75 on different lysates with Rabbit anti-Grp75 antibody (R1510-38) at 1/1,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: K-562 cell lysate (20 µg/Lane) Lane 3: NIH/3T3 cell lysate (20 µg/Lane) Lane 4: PC-12 cell lysate (20 µg/Lane) Lane 5: Mouse liver tissue lysate (40 µg/Lane) Lane 6: Mouse kidney tissue lysate (40 µg/Lane) Lane 7: Rat liver tissue lysate (40 µg/Lane) Predicted band size: 74 kDa Observed band size: 74 kDa Exposure time: 2 seconds; ECL: K1801; 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 (R1510-38) 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. |
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Fig2:
Immunocytochemistry analysis of HeLa cells labeling Grp75 with Rabbit anti-Grp75 antibody (R1510-38) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Grp75 antibody (R1510-38) at 1/200 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 (M1305-2, 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. |
Fig3:
Immunocytochemistry analysis of NIH/3T3 cells labeling Grp75 with Rabbit anti-Grp75 antibody (R1510-38) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Grp75 antibody (R1510-38) 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 (M1305-2, 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-Grp75 antibody (R1510-38) at 1/200 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 (R1510-38) at 1/200 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Grp75 antibody (R1510-38) at 1/200 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 (R1510-38) at 1/200 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. |
Fig6:
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Grp75 antibody (R1510-38) at 1/200 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 (R1510-38) at 1/200 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. |
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Fig7:
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Grp75 antibody (R1510-38) at 1/200 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 (R1510-38) at 1/200 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. |
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Fig8: Flow cytometric analysis of Grp75 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (R1510-38, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red). |