Validated using a knockout cell line

Anti-Calpain S1 antibody [EPR3324] (ab92333)


  • Product name
    Anti-Calpain S1 antibody [EPR3324]
    See all Calpain S1 primary antibodies
  • Description
    Rabbit monoclonal [EPR3324] to Calpain S1
  • Host species
  • Tested applications
    Suitable for: WB, IHC-P, ICC, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Chinese hamster
  • Immunogen

    Synthetic peptide (Human)

  • Positive control
    • WB: fetal brain or fetal spleen tissue lysate; T47D or 293T cell lysate. IHC: Human kidney tissue.
  • 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

    This product is a recombinant rabbit monoclonal antibody.



Our Abpromise guarantee covers the use of ab92333 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
Flow Cyt
  • Application notes
    Flow Cyt: 1/50.
    ICC: 1/100 - 1/250.
    IHC-P: 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. The use of an HRP/AP polymerized antibody will give a stronger signal.

    WB: 1/1000 - 1/5000. Predicted molecular weight: 28 kDa.

    Is unsuitable for IP.

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      Regulatory subunit of the calcium-regulated non-lysosomal thiol-protease which catalyzes limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.
    • Sequence similarities
      Contains 5 EF-hand domains.
    • Domain
      The contact of the 5th EF-hand domain from each monomer allows the formation of the homodimer and also appears to mediate the contact between the large catalytic subunit and small regulatory subunit for the formation of the heterodimer.
      EF-hand domains are paired. EF-hand 1 is paired with EF-hand 2 and EF-hand 3 is paired with EF-hand 4. The fifth EF-hand domain, left unpaired, does not bind the calcium but is responsible of the dimerization by EF-embrace. The first four EF-hand domains bind calcium, however it is not sure if the binding of EF-hand 4 to calcium is physiologically relevant.
    • Cellular localization
      Cytoplasm. Cell membrane. Translocates to the plasma membrane upon calcium binding.
    • Information by UniProt
    • Database links
    • Alternative names
      • 30K antibody
      • Calcium activated neutral proteinase antibody
      • Calcium activated neutral proteinase small subunit antibody
      • Calcium dependent protease small subunit 1 antibody
      • Calcium dependent protease small subunit antibody
      • Calcium-activated neutral proteinase small subunit antibody
      • Calcium-dependent protease small subunit 1 antibody
      • Calcium-dependent protease small subunit antibody
      • Calpain 4 antibody
      • Calpain 4 small subunit (30K) antibody
      • Calpain 4 small subunit antibody
      • Calpain regulatory subunit antibody
      • Calpain small polypeptide antibody
      • Calpain small subunit 1 antibody
      • Calpain4 antibody
      • CalpainS1 antibody
      • CANP antibody
      • CANP small subunit antibody
      • CANPS antibody
      • CAPN 4 antibody
      • CAPN S1 antibody
      • CAPN4 antibody
      • CAPNS 1 antibody
      • CAPNS antibody
      • Capns1 antibody
      • CDPS antibody
      • CPNS1_HUMAN antibody
      • CSS 1 antibody
      • CSS1 antibody
      see all


    • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: CALPAIN S1 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: A431 whole cell lysate (20 µg)
      Lane 4: HEK293 whole cell lysate (20 µg)


      Lanes 1 - 4: Merged signal (red and green). Green - ab92333 observed at 28 kDa. Red - loading control, ab8245, observed at 37 kDa.


      ab92333 was shown to recognize CALPAIN S1 when CALPAIN S1 knockout samples were used, along with additional cross-reactive bands. Wild-type and CALPAIN S1 knockout samples were subjected to SDS-PAGE. Ab92333 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 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.

    • All lanes : Anti-Calpain S1 antibody [EPR3324] (ab92333) at 1/2000 dilution

      Lane 1 : T47D
      cell lysate
      Lane 2 : fetal brain lysate
      Lane 3 : fetal spleen lysate
      Lane 4 : 293T
      cell lysate

      Lysates/proteins at 10 µg per lane.

      All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

      Predicted band size: 28 kDa
      Observed band size: 28 kDa

    • Immunohistochemistry staining of Calpain S1 in formalin-fixed, paraffin-embedded Human kidney tissue using 1/100 ab92333.
    • Overlay histogram showing HeLa cells stained with ab92333 (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 (ab92333, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.


    This product has been referenced in:
    • Guo Y  et al. Remarkable Reduction of MAP2 in the Brains of Scrapie-Infected Rodents and Human Prion Disease Possibly Correlated with the Increase of Calpain. PLoS One 7:e30163 (2012). WB ; Chinese hamster . Read more (PubMed: 22272295) »

    See 1 Publication for this product

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