Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)

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Western blot - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
  • Immunoprecipitation - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
  • Flow Cytometry - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
  • Immunocytochemistry/ Immunofluorescence - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
  • Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20681] to CYR61/CCN1 - BSA and Azide free
  • Suitable for: WB, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free
    See all CYR61/CCN1 primary antibodies

  • Description

    Rabbit monoclonal [EPR20681] to CYR61/CCN1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human CYR61/CCN1 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: O00622

  • Positive control

    • ICC/IF: MDA-MB-231 cells. WB: MDA-MB-231, MCF7 and HeLa cell lysates. Flow Cyt: MDA-MB-231 and MCF7 cells. IP: Saos-2 cell lysate.

  • General notes

    Ab233097 is the carrier-free version of ab230947. 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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab233097 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab233097 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration. Predicted molecular weight: 42 kDa.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

  • Function

    Promotes cell proliferation, chemotaxis, angiogenesis and cell adhesion. Appears to play a role in wound healing by up-regulating, in skin fibroblasts, the expression of a number of genes involved in angiogenesis, inflammation and matrix remodeling including VEGA-A, VEGA-C, MMP1, MMP3, TIMP1, uPA, PAI-1 and integrins alpha-3 and alpha-5. CYR61-mediated gene regulation is dependent on heparin-binding. Down-regulates the expression of alpha-1 and alpha-2 subunits of collagen type-1. Promotes cell adhesion and adhesive signaling through integrin alpha-6/beta-1, cell migration through integrin alpha-v/beta-5 and cell proliferation through integrin alpha-v/beta-3.

  • Sequence similarities

    Belongs to the CCN family.
    Contains 1 CTCK (C-terminal cystine knot-like) domain.
    Contains 1 IGFBP N-terminal domain.
    Contains 1 TSP type-1 domain.
    Contains 1 VWFC domain.

  • Cellular localization

    Secreted.
  • Information by UniProt

  • Database links

    • Alternative names

      • 3CH61 antibody
      • CCN 1 antibody
      • CCN family member 1 antibody
      • CCN1 antibody
      • CYR 61 antibody
      • Cyr61 antibody
      • CYR61 protein antibody
      • CYR61_HUMAN antibody
      • Cysteine rich angiogenic inducer 61 antibody
      • Cysteine rich heparin binding protein 61 antibody
      • Cysteine-rich angiogenic inducer 61 antibody
      • GIG 1 antibody
      • GIG1 antibody
      • IBP-10 antibody
      • IGF-binding protein 10 antibody
      • Igfbp 10 antibody
      • IGFBP-10 antibody
      • Igfbp10 antibody
      • Insulin like growth factor binding protein 10 antibody
      • Insulin-like growth factor-binding protein 10 antibody
      • Protein CYR61 antibody
      • Protein GIG1 antibody
      see all

    Images

    • Western blot - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
      Western blot - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
      All lanes : Anti-CYR61/CCN1 antibody [EPR20681] (ab230947) at 1/500 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : CYR61 knockout HeLa cell lysate
      Lane 3 : MDA-MB-231 cell lysate
      Lane 4 : MCF7 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 42 kDa
      Observed band size: 47 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 (ab230947).

      Lanes 1-4: Merged signal (red and green). Green - ab230947 observed at 47 kDa. Red - loading control ab8245 observed at 36 kDa. 

       ab230947 Anti-CYR61/CCN1 antibody [EPR20681] was shown to specifically react with CYR61/CCN1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265288 (knockout cell lysate ab257406) was used.  Wild-type and CYR61/CCN1 knockout samples were subjected to SDS-PAGE.  ab230947 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 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-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
      Immunoprecipitation - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)

      CYR61/CCN1 was immunoprecipitated from 0.35 mg Saos-2 (human osteosarcoma epithelial) whole cell lysate with ab230947 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230947 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
      Lane 1: Saos-2 (human osteosarcoma epithelial) whole cell lysate 10 µg (Input).
      Lane 2: ab230947 IP in Saos-2 whole cell lysate (+).
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab230947 in Saos-2 whole cell lysate (-).
      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 8 seconds.

      The molecular mass observed is consistent with that of the full-length protein (42 kDa) (PMID: 23798676).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab230947).

    • Flow Cytometry - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
      Flow Cytometry - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell, Left) / MDA-MB-231 (human breast adenocarcinoma epithelial cell, Right) cell line labeling CYR61/CCN1 with ab230947 at 1/60 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

      Negative control: MCF7 (PMID: 11059746).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab230947).

    • Immunocytochemistry/ Immunofluorescence - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
      Immunocytochemistry/ Immunofluorescence - Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDA-MB-231 (human breast adenocarcinoma epithelial cell) cells labeling CYR61/CCN1 with ab230947 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in the MDA-MB-231 cell line. Negative control: MCF7 (PMID: 11059746). The nuclear counter stain is DAPI (blue).
      Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
      The negative control is the secondary antibody only.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab230947).

    • Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)
      Anti-CYR61/CCN1 antibody [EPR20681] - BSA and Azide free (ab233097)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab233097? Please let us know so that we can cite the reference in this datasheet.

    ab233097 has not yet been referenced specifically in any publications.

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