RhoA Recombinant Rabbit Monoclonal Antibody [SN0612]
cat.: ET1611-10
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Tissue, FC, IHC-P, IF-Cell
Clonality: Monoclonal
Clone number: SN0612
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
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: 22 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human aa 144-193 / 193.
Positive control: HUVEC cell lysate, THP-1 cell lysate, HeLa cell lysate, SiHa cell lysate, mouse brain tissue lysate, rat brain tissue lysate, human brain tissue lysate, PC-12, human liver cancer tissue, mouse brain tissue, Hela.
Subcellular location: Cell membrane, Cytoplasm, Cleavage furrow, Midbody, Cell projection.
Recommended Dilutions:
  WB
  IF-Tissue
  FC
  IHC-P
  IF-Cell
1:1,000-1:2,000
1:50-1:200
1:50-1:100
1:50-1:200
1:100
Uniprot #: SwissProt: P61586 Human | Q9QUI0 Mouse | P61589 Rat
Alternative names: Aplysia ras related homolog 12 ARH12 ARHA H 12 H12 Oncogene RHO H12 Ras homolog family member A Ras homolog gene family member A Rho A Rho cDNA clone 12 RHO H12 RHO12 RHOA RHOA_HUMAN RHOH12 Small GTP binding protein Rho A Transforming protein Rho A Transforming protein RhoA
Images
ET1611-10_1.jpg Fig1: Western blot analysis of RhoA on different lysates with Rabbit anti-RhoA antibody (ET1611-10) at 1/1,000 dilution.

Lane 1: HUVEC cell lysate (20 µg/Lane)
Lane 2: THP-1 cell lysate (20 µg/Lane)
Lane 3: HeLa cell lysate (20 µg/Lane)
Lane 4: SiHa cell lysate (20 µg/Lane)
Lane 5: Mouse brain tissue lysate (40 µg/Lane)
Lane 6: Rat brain tissue lysate (40 µg/Lane)
Lane 7: Human brain tissue lysate (40 µg/Lane)

Predicted band size: 22 kDa
Observed band size: 22 kDa

Exposure time: 1 minute;

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 (ET1611-10) 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 1:100,000 dilution was used for 1 hour at room temperature.
ET1611-10_2.jpg Fig2: Immunocytochemistry analysis of PC-12 cells labeling RhoA with Rabbit anti-RhoA antibody (ET1611-10) at 1/100 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-RhoA antibody (ET1611-10) 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 (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.
ET1611-10_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-RhoA antibody (ET1611-10) at 1/50 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 (ET1611-10) at 1/50 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.
ET1611-10_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-RhoA antibody (ET1611-10) 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 (ET1611-10) 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.
ET1611-10_5.jpg Fig5: Flow cytometric analysis of RhoA was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1611-10, 1/50) (red). 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 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.