RhoA/B/C Antibody

RhoA/B/C Antibody is a non-conjugated and Rabbit origined polyclonal antibody about 22 kDa, targeting to RhoA/B/C. It can be used for WB,IHC-P,ICC/IF,FC assays with tag free, in the background of Human, Mouse, Rat.

RhoA Protein is a small GTPase crucial for orchestrating diverse cellular responses, cycling between an active GTP-bound state and an inactive GDP-bound state. Primarily associated with the organization of the cytoskeleton, the active form of RhoA binds to various effector proteins, influencing cellular processes such as cytoskeletal dynamics, cell migration, and cell cycle progression. It plays a pivotal role in regulating signal transduction pathways that connect plasma membrane receptors to the assembly of focal adhesions and actin stress fibers. Additionally, RhoA is indispensable for microtubule-dependent signals required for myosin contractile ring formation during cell cycle cytokinesis, contributing to cleavage furrow formation. Essential for apical junction formation in keratinocyte cell-cell adhesion, RhoA also participates in the MEMO1-RHOA-DIAPH1 signaling pathway, influencing ERBB2-dependent microtubule stabilization. It further modulates potassium channel activity, regulates guanine nucleotide exchange factors, and acts as a target for the yopT cysteine peptidase from Yersinia pestis during microbial infection.

Free

P61586 / P62745 / P08134

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

affinity purified

Non-conjugated

Unmodified

AB_3102989

Signal Transduction

Primary Antibody; Rabbit Polyclonal Antibody

Polyclonal

Rabbit

Human, Mouse, Rat

WB: 1:500-1:1,000 ;IHC: 1:50-1:100 ;IF: 1:50-1:200 ;FC: 1:50-1:100

• 
  Western blot analysis of extracts from NIH/3T3(lane 2(20ug) ,HepG2(lane 3(20ug) and Hela(lane 4(20ug) using RhoA/B/C Antibody (HY-P80882) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of HepG2 cells labeling RhoA/B/C with RhoA/B/C Antibody (HY-P80882) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RhoA/B/C Antibody (HY-P80882) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of NIH3T3 cells labeling RhoA/B/C with RhoA/B/C Antibody (HY-P80882) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with RhoA/B/C Antibody (HY-P80882) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded rat pancreas tissue using Collagen VI alpha 123 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. 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.
• 
  Immunohistochemical analysis of paraffin-embedded rat pancreas tissue using Collagen VI alpha 123 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. 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.

WB, IHC-P, ICC/IF, FC

Liquid

Supplied in 50mM Tris-Glycine(pH 7.4), 0.15M NaCl, 40%Glycerol, 0.01% sodium azide and 0.05% BSA.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human Rho A + B + C.The exact sequence is proprietary to MCE.