Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP786Y] to ROCK2 + ROCK1 - BSA and Azide free
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free

  • Description

    Rabbit monoclonal [EP786Y] to ROCK2 + ROCK1 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody recognizes both the cleaved C-terminus of ROCK 1 (30 kDa) and full length protein (158 kDa). The immunogen used for this product shares 83% homology with ROCK2 and has been shown to bind recombinant human ROCK2, please see western blot images below.

  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details

  • Species reactivity

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

  • Immunogen

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

  • Positive control

    • WB: Untreated, Calyculin A treated and Camptothecin treated HeLa cell lysates. Jurkat, Ramos, PC-12 and RAW264.7 cell lysates. IHC-P: Human adenocarcinoma of the colon and thyroid gland carcinoma tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: HeLa cell lysate.

  • General notes

    Ab219587 is the carrier-free version of ab45171. 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

    ab219587 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

  • Immunoprecipitation - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
    Immunoprecipitation - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)

    ab45171 (purified) at 1/40 immunoprecipitating ROCK2 + ROCK1 in HeLa cell lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).

  • Immunocytochemistry/ Immunofluorescence - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
    Immunocytochemistry/ Immunofluorescence - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ROCK2 + ROCK1 with purified ab45171 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150078, an Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/50) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of the colon tissue labelling ROCK2 +  ROCK1 with purified ab45171 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587) This image is courtesy of an anonymous Abreview.

    Unpurified ab45171 staining ROCK2 + ROCK1 in mouse embyro 18dpc tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 2 hours at room temperature; antigen retrieval was by heat mediation in Tris pH 9. Samples were incubated with primary antibody (1/100 in 1% BSA + 1% FBS in TBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal 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 (ab45171).

  • Flow Cytometry - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
    Flow Cytometry - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)

    Overlay histogram showing HeLa cells stained with unpurified ab45171 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab45171, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).

  • Western blot - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
    Western blot - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
    All lanes : Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171) at 1/1000 dilution

    Lane 1 : Recombinant Human ROCK1 protein (aa 1114 to 1354) (30 kDa)
    Lane 2 : Recombinant Human ROCK2 protein (aa 1132 to 1388) (30 kDa)

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 158 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab45171, the same antibody clone in a different buffer formulation.

    Loading control: Anti-6X His tag® antibody [EPR20547] (ab213204)

    Blocking buffer and concentration: 5% NFDM/TBST

    Exposure Times:

    Lane 1: 3 seconds

    Lane 2: 7 seconds

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling ROCK2 + ROCK1 with unpurified ab45171.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
    Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)

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