Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18299] to Rho A + B + C - BSA and Azide free
  • Suitable for: ICC, IHC-P, Flow Cyt, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free
    See all Rho A + B + C primary antibodies

  • Description

    Rabbit monoclonal [EPR18299] to Rho A + B + C - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human RhoA, RhoB and RhoC full length proteins; HepG2, HeLa, Jurkat, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal brain and fetal kidney lysates; Mouse and rat kidney and brain lysates. IHC-P: Human stomach and squamous cell carcinoma of lung, Mouse muscle and Rat colon tissues. ICC/IF: HepG2 and NIH/3T3 cells. Flow Cyt: Jurkat cells.

  • General notes

    Ab221161 is the carrier-free version of ab188103. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab221161 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    All lanes : Anti-Rho A + B + C antibody [EPR18299] (ab188103) at 1/20000 dilution

    Lane 1 : Human RhoA full length protein
    Lane 2 : Human RhoB full length protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 24 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    This data was developed using ab188103, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Human RhoA full length protein (ab101594) contains aa1-193 with His-Tag® N-Terminus; Human RhoB full length protein (ab107139) contains aa1-193 with His-Tag®; N-Terminus.

  • Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Anti-Rho A + B + C antibody [EPR18299] (ab188103) at 1/500 dilution + Human RhoC full length protein at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 24 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    This data was developed using ab188103, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Human RhoC full length protein (ab98085) contains aa1-190 with His-Tag® N-Terminus.

  • Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    All lanes : Anti-Rho A + B + C antibody [EPR18299] (ab188103) at 1/2000 dilution

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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


    Exposure time: 1 minute


    This data was developed using ab188103, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    All lanes : Anti-Rho A + B + C antibody [EPR18299] (ab188103) at 1/2000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

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


    Exposure time: 1 minute


    This data was developed using ab188103, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Western blot - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    All lanes : Anti-Rho A + B + C antibody [EPR18299] (ab188103) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Rat kidney lysate
    Lane 5 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 6 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 7 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 8 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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


    Exposure time: 1 minute


    This data was developed using ab188103, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)

    This data was developed using ab188103, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling Rho A + B + C with ab188103 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic and nuclear staining on epithelial cells of Human stomach is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)

    This data was developed using ab188103, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of lung tissue labeling Rho A + B + C with ab188103 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic and nuclear staining on cancer cells of squamous cell carcinoma of lung is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    This data was developed using ab188103, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse muscle tissue labeling Rho A + B + C with ab188103 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic and nuclear staining on mouse muscle is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    This data was developed using ab188103, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Rho A + B + C with ab188103 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic and nuclear staining on epithelial cells of Rat colon is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunocytochemistry - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Immunocytochemistry - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    This data was developed using ab188103, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling Rho A + B + C with ab188103 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing both nuclear and cytoplasmic staining on HepG2 cells. 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 1/500 dilution (red).
    The negative controls are as follows;
    -ve control 1: ab188103 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
  • Immunocytochemistry - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Immunocytochemistry - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    This data was developed using ab188103, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling to RhoA + RhoB + RhoC with ab188103 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing both nuclear and cytoplasmic staining on NIH/3T3 cells. 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 1/500 dilution (red).
    The negative controls are as follows;
    -ve control 1: ab188103 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
  • Flow Cytometry - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Flow Cytometry - Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    This data was developed using ab188103, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Rho A + B + C with ab188103 at 1/300 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
  • Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)
    Anti-Rho A + B + C antibody [EPR18299] - BSA and Azide free (ab221161)

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