Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR638Y] to ROCK1 - BSA and Azide free
  • Suitable for: WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free
    See all ROCK1 primary antibodies

  • Description

    Rabbit monoclonal [EPR638Y] to ROCK1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HeLa cell lysate, HEK293 cell lysate IHC: Human lymphoma tissue. IP: HeLa whole cell lysate.

  • General notes

    Ab230799 is the carrier-free version of ab134181. 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

    ab230799 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Images

  • Western blot - Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)
    Western blot - Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)

    This WB data was generated using the same anti-ROCK1 antibody clone, EPR638Y, in a different buffer formulation (cat# ab134181).

    Lane 1: Wild type HAP1 whole cell lysate (40 µg)
    Lane 2: ROCK1 knockout HAP1 whole cell lysate (40 µg)
    Lane 3: HEK293 whole cell lysate (20 µg)
    Lane 4: HeLa whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab134181 observed at 165 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab134181 was shown to specifically react with ROCK1 when ROCK1 knockout samples were used. Wild-type and ROCK1 knockout samples were subjected to SDS-PAGE. Ab134181 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)

    Immunohistochemical analysis of paraffin embedded Human lymphoma tissue labelling ROCK1 with ab134181 at a dilution of 1/50.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134181).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)
    Western blot - Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)
    All lanes : Anti-ROCK1 antibody [EPR638Y] (ab134181) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ROCK1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 158 kDa
    Observed band size: 160 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 (ab134181).

    Lanes 1- 2: Merged signal (red and green). Green - ab134181 observed at 160 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab134181 was shown to react with ROCK1 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264780 (knockout cell lysate ab257642) lane below 160kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and ROCK1 knockout HeLa 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. ab134181 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 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.

  • Immunoprecipitation - Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)
    Immunoprecipitation - Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)

    Purified ab134181 at 1/120 dilution (2µg) immunoprecipitating ROCK1 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab134181 + HeLa whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab134181 in HeLa whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 158 kDa
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134181).

  • Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)
    Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)

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