Recombinant
RabMAb

Anti-Calponin antibody [EP798Y] (ab46794)

Overview

  • Product name
    Anti-Calponin antibody [EP798Y]
    See all Calponin primary antibodies
  • Description
    Rabbit monoclonal [EP798Y] to Calponin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human, Pig
  • Immunogen

    Synthetic peptide corresponding to Human Calponin aa 250-350 (C terminal).

  • Positive control
    • Human bladder lysate; HeLa cells; Human smooth muscle tissue. Mouse bladder lysates, rat bladder lysates, pig heart lysates. Human lung carcinoma, mouse cardiac muscle, C2C12.
  • General notes

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab46794 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 1/5000. Detects a band of approximately 34 kDa.

For unpurified use at 1/20,000.

IHC-P 1/1000.

For unpurified use at 1/100 - 1/250.

ICC/IF 1/500.

For unpurified use at 1/100 - 1/250. 

  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Function
      Thin filament-associated protein that is implicated in the regulation and modulation of smooth muscle contraction. It is capable of binding to actin, calmodulin, troponin C and tropomyosin. The interaction of calponin with actin inhibits the actomyosin Mg-ATPase activity.
    • Tissue specificity
      Smooth muscle, and tissues containing significant amounts of smooth muscle.
    • Sequence similarities
      Belongs to the calponin family.
      Contains 3 calponin-like repeats.
      Contains 1 CH (calponin-homology) domain.
    • Information by UniProt
    • Database links
    • Alternative names
      • Basic calponin antibody
      • Calponin 1 antibody
      • Calponin 1 basic smooth muscle antibody
      • Calponin H1 antibody
      • Calponin H1 smooth muscle antibody
      • Calponin-1 antibody
      • Calponin1 antibody
      • Calponins basic antibody
      • CNN 1 antibody
      • Cnn1 antibody
      • CNN1_HUMAN antibody
      • Epididymis secretory protein Li 14 antibody
      • HEL S 14 antibody
      • Sm Calp antibody
      • SMCC antibody
      • smooth muscle antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence analysis of C2C12 (Mouse myoblasts myoblast) cells labeling Calponin with Purified ab46794 at 1:500 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor ® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor ® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat lung tissue sections labeling Calponin with Purified ab46794 at 1:1000 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
    • All lanes : Anti-Calponin antibody [EP798Y] (ab46794) at 1/50000 dilution (purified)

      Lane 1 : Human bladder lysates
      Lane 2 : Mouse bladder lysates
      Lane 3 : Rat bladder lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution


      Blocking and diluting buffer: 5% NFDM/TBST.

    • Anti-Calponin antibody [EP798Y] (ab46794) at 1/5000 dilution (purified) + Pig heart lysates at 15 µg

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


      Blocking and diluting buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cardiac muscle tissue sections labeling Calponin with Purified ab46794 at 1:1000 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling Calponin with Purified ab46794 at 1:1000 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
    • Unpurified ab46794 staining Calponin in porcine aortic smooth muscle cells by Immunocytochemistry/ Immunofluorescence. The cells were paraformaldehyde fixed, permeabilised in 0.1% Triton X-100. Samples were then incubated with primary antibody at 1/50 for 1 hour at 25°C. The secondary antibody used was ab6717 Goat polyclonal to Rabbit IgG - H&L (FITC) (green) used at a 1/400 dilution.

      See Abreview

    • Immunohistochemical staining of paraffin-embedded human smooth muscle using unpurified ab46794 at 1/100 dilution

    • ICC/IF image of unpurified ab46794 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab46794, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Anti-Calponin antibody [EP798Y] (ab46794) at 1/20000 dilution (unpurified) + Human bladder lysate at 10 µg

      Secondary
      Goat anti-rabbit HRP at 1/2000 dilution

      Observed band size: 34 kDa (why is the actual band size different from the predicted?)

    • All lanes : Anti-Calponin antibody [EP798Y] (ab46794) at 1/20000 dilution (unpurified)

      All lanes : Whole tissue lysate prepared from bovine lymph vessels

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP conjugated goat anti-rabbit polyclonal at 1/5000 dilution

      Developed using the ECL technique.

      Observed band size: 34 kDa (why is the actual band size different from the predicted?)


      Exposure time: 1 minute

      See Abreview

    • Unpurified ab46794 showing positive staining in Normal kidney vessels tissue.

    • Unpurified ab46794 showing positive staining in Normal lung vessel tissue.

    • Unpurified ab46794 showing positive staining in Normal tonsil vessel tissue.

    • Unpurified ab46794 showing positive staining in Normal uterus tissue.

    • Unpurified ab46794 showing negative staining in Skeletal muscle tissue.

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    See all 83 Publications for this product

    Customer reviews and Q&As

    Application
    Immunocytochemistry
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1.5% · Temperature: 25°C
    Sample
    Mouse Cultured Cells (thoracic aortic smooth muscle cells)
    Specification
    thoracic aortic smooth muscle cells
    Permeabilization
    Yes - 0.25% TritonX-100
    Fixative
    Paraformaldehyde
    Username

    Mr. Mc Shen

    Verified customer

    Submitted Dec 15 2014

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Sheep Tissue sections (Artery)
    Permeabilization
    Yes - 0.1% Triton X-100, 10 min
    Specification
    Artery
    Blocking step
    Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative
    Paraformaldehyde
    Username

    Ms. Chelsea Stowell

    Verified customer

    Submitted Mar 07 2018

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Smooth muscle cells)
    Permeabilization
    Yes - 0.15% Triton-X-100
    Specification
    Smooth muscle cells
    Blocking step
    BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Mar 05 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Prostate)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate, pH 6
    Permeabilization
    No
    Specification
    Prostate
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Dec 06 2017

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Western blot
    Sample
    Cow Tissue lysate - whole (Kidney)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Kidney
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Sep 22 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Breast)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA pH9
    Permeabilization
    Yes - TBST-5 min
    Specification
    Breast
    Blocking step
    (agent) for 10 minute(s) · Concentration: 0%
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jun 10 2016

    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (vascular smooth muscle cell)
    Specification
    vascular smooth muscle cell
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Username

    Abcam user community

    Verified customer

    Submitted Jul 07 2014

    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (11% SDS-PAGE)
    Sample
    Human Cell lysate - whole cell (HUVECells and 5ng/ml TNF-a for 24hr)
    Specification
    HUVECells and 5ng/ml TNF-a for 24hr
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Aug 09 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (smooth muscle cells)
    Specification
    smooth muscle cells
    Fixative
    Acetone
    Permeabilization
    Yes - acetone permeabilizes as well
    Blocking step
    Serum as blocking agent for 40 minute(s) · Concentration: 25% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 18 2012

    Thank you for contacting us.

    In order to redeem the free antibody, please use the code that was provided to you (xxx) on your next order that you place with Abcam (before the expiration date: xxx).

    As per instructions:
    The code wi...

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