Overview

  • Product name

    Anti-Calpastatin antibody [SP82]
    See all Calpastatin primary antibodies
  • Description

    Rabbit monoclonal [SP82] to Calpastatin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Calpastatin aa 550-650 (C terminal). The exact sequence is proprietary.
    Database link: P20810

  • Positive control

    • IHC-P: Human breast carcinoma tissue; FC: HaCaT and HeLa cells; ICC: HaCaT cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab101684 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
ICC/IF Use at an assay dependent concentration.
IHC-P 1/100.

Incubate for 30 minutes at RT. Antigen retrieval is recommended: boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.

Flow Cyt Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    Specific inhibition of calpain (calcium-dependent cysteine protease). Plays a key role in postmortem tenderization of meat and have been proposed to be involved in muscle protein degradation in living tissue.
  • Sequence similarities

    Belongs to the protease inhibitor I27 (calpastatin) family.
  • Domain

    Has four inhibitory domains.
  • Post-translational
    modifications

    The N-terminus is blocked.
  • Information by UniProt
  • Database links

  • Alternative names

    • BS 17 antibody
    • Calpain inhibitor antibody
    • Calpastatin antibody
    • Cast antibody
    • Heart type calpastatin antibody
    • ICAL_HUMAN antibody
    • MGC9402 antibody
    • Sperm BS 17 component antibody
    • Sperm BS-17 component antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling Calpastatin with Purified ab101684 at 1/100 dilution (0.59 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunocytochemistry/ Immunofluorescence analysis of HaCaT (human skin keratinocyte) cells labeling Calpastatin with purified ab101684. Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow cytometry analysis of HaCaT (human skin keratinocyte) labeling Calpastatin with purified ab101684 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (black). Unlableled control - Unlabelled cells (blue).

  • Immunohistochemical analysis of Calpastatin in formalin-fixed, paraffin-embedded Human breast carcinoma using ab101684 at 1/200 dilution.
  • Overlay histogram showing HeLa cells stained with ab101684 (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 (ab101684) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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.

References

ab101684 has not yet been referenced specifically in any publications.

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