R1510-39

Grp75 Rabbit Polyclonal Antibody

cat.: R1510-39

Product Type:	Rabbit polyclonal IgG, primary antibodies
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 human Grp75 aa 114-157.
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	1:1,000-1:2,000 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

Images

Fig1: Western blot analysis of Grp75 on different lysates with Rabbit anti-Grp75 antibody (R1510-39) 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 (R1510-39) 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.

Fig2: Immunocytochemistry analysis of HeLa cells labeling Grp75 with Rabbit anti-Grp75 antibody (R1510-39) 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 (R1510-39) 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 (R1510-39) 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 (R1510-39) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-Grp75 antibody (R1510-39) 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 (R1510-39) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Grp75 antibody (R1510-39) 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 (R1510-39) 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 (R1510-39) 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 (R1510-39) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Grp75 antibody (R1510-39) 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 (R1510-39) 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.
	Fig8: Flow cytometric analysis of Grp75 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (R1510-39, 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).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.