Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18134] to RhoA - Low endotoxin, Azide free
  • Suitable for: WB, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free
    See all RhoA primary antibodies

  • Description

    Rabbit monoclonal [EPR18134] to RhoA - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human RhoA full length protein; HeLa, HEK293 C6, Raw264.7 and NIH 3T3 cell lysates, human fetal brain and fetal kidney lysates, mouse brain, kidney and spleen lysates, rat brain, kidney and spleen lysates. ICC/IF: Jurkat and K562 cells. Flow Cyt: HeLa cells.

  • General notes

    ab219371 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/5000 dilution

    Lanes 1 & 3 : Wild-type HEK-293T cell lysate
    Lanes 2 & 4 : RHOA knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 22 kDa
    Observed band size: 21 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab187027).

    Lanes 1- 4: Merged signal (red and green). Green - ab187027 observed at 21 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab187027 was shown to react with RhoA in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266592 (knockout cell lysate ab257637) was used. Wild-type HEK-293T and RHOA knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab187027 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling RhoA with ab187027 at 1/150 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Jurkat cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab187027 at 1/150 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187027).

  • Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Western blot - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    All lanes : Anti-RhoA antibody [EPR18134] (ab187027) at 1/1000 dilution

    Lane 1 : Wild-type Hek293T whole cell lysate
    Lane 2 : RHOA knockout Hek293T whole cell lysate
    Lane 3 : Jurkat whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 22 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab187027 observed at 22 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab187027 was shown to specifically react with RhoA in wild-type Hek293T cells as signal was lost in RHOA knockout cells. Wild-type and RHOA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab187027 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187027).

  • Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Immunocytochemistry/ Immunofluorescence - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling RhoA with ab187027 at 1/150 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on K562 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab187027 at 1/150 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187027).

  • Flow Cytometry - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Flow Cytometry - Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling RhoA with ab187027 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and a unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187027).

  • Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)
    Anti-RhoA antibody [EPR18134] - Low endotoxin, Azide free (ab219371)

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