RhoGAP Recombinant Rabbit Monoclonal Antibody [JE53-53]
cat.: ET7110-65
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Monoclonal
Clone number: JE53-53
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 172 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human ARHGAP5 aa 1,130-1,320 / 1,502.
Positive control: MCF7 cell lysate, C2C12 cell lysate, C6 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, human testis tissue, mouse testis tissue, rat testis tissue, MCF7, C2C12, PC-12.
Subcellular location: Cell membrane, cytoplasm.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
1:1,000-1:5,000
1:1,000
1:100
1:1,000
Uniprot #: SwissProt: Q13017 Human | P97393 Mouse
Entrez Gene: 299012 Rat
Alternative names: ARHGAP 5 ARHGAP5 p190 B p190 RhoGAP Rho GTPase activating protein 5 RhoGAP 5 RhoGAP RhoGAP5
Images
ET7110-65_1.jpg Fig1: Western blot analysis of RhoGAP on different lysates with Rabbit anti-RhoGAP antibody (ET7110-65) at 1/5,000 dilution.

Lane 1: MCF7 cell lysate
Lane 2: C2C12 cell lysate
Lane 3: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 172 kDa
Observed band size: 172 kDa

Exposure time: 30 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 (ET7110-65) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET7110-65_2.jpg Fig2: Western blot analysis of RhoGAP on different lysates with Rabbit anti-RhoGAP antibody (ET7110-65) at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate
Lane 2: PC-12 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 172 kDa
Observed band size: 172 kDa

Exposure time: 3 minutes; 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 (ET7110-65) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET7110-65_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-RhoGAP antibody (ET7110-65) 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 (ET7110-65) 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.
ET7110-65_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-RhoGAP antibody (ET7110-65) 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 (ET7110-65) 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.
ET7110-65_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-RhoGAP antibody (ET7110-65) 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 (ET7110-65) 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.
ET7110-65_6.jpg Fig6: Immunocytochemistry analysis of MCF7 cells labeling RhoGAP with Rabbit anti-RhoGAP antibody (ET7110-65) at 1/100 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-RhoGAP antibody (ET7110-65) at 1/100 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.
ET7110-65_7.jpg Fig7: Immunocytochemistry analysis of C2C12 cells labeling RhoGAP with Rabbit anti-RhoGAP antibody (ET7110-65) at 1/100 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-RhoGAP antibody (ET7110-65) at 1/100 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.
ET7110-65_8.jpg Fig8: Immunocytochemistry analysis of PC-12 cells labeling RhoGAP with Rabbit anti-RhoGAP antibody (ET7110-65) at 1/100 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-RhoGAP antibody (ET7110-65) at 1/100 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.
ET7110-65_9.jpg Fig9: Flow cytometric analysis of MCF7 cells labeling RhoGAP.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET7110-65, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET7110-65_10.jpg Fig10: Flow cytometric analysis of C2C12 cells labeling RhoGAP.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET7110-65, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET7110-65_11.jpg Fig11: Flow cytometric analysis of PC-12 cells labeling RhoGAP.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET7110-65, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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