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Signal Transduction Metabolism Amino Acids
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)

  • Datasheet
  • SDS
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Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
  • Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
  • Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3323] to Calpain small subunit 1
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human

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Overview

  • Product name

    Anti-Calpain small subunit 1 antibody [EPR3323]
    See all Calpain small subunit 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR3323] to Calpain small subunit 1
  • Host species

    Rabbit
  • Specificity

    Detects Calpain small subunit 1.
  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human Calpain small subunit 1 aa 50-150. The exact sequence is proprietary.
    Database link: P04632

  • Positive control

    • T47-D, 293T, A431 and Human spleen lysates. Human kidney tissue.
  • General notes

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR3323
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Amino Acids
    • Signal Transduction
    • Signaling Pathway
    • Calcium Signaling
    • Calpain
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Cysteine protease
    • Calpains
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Amino acid metabolism

Associated products

  • Alternative Versions

    • Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • A-431 membrane extract lysate (ab30134)
  • Recombinant Protein

    • Recombinant Human Calpain small subunit 1 protein (ab180298)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab92356 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Human
WB
Human
All applications
Rat
Application Abreviews Notes
WB
1/1000 - 1/2000. Predicted molecular weight: 28 kDa.
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 is recommended as these have been shown to give superior staining.

Notes
WB
1/1000 - 1/2000. Predicted molecular weight: 28 kDa.
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 is recommended as these have been shown to give superior staining.

  • Application notes
    Is unsuitable for IP.
  • 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.
    • Target information above from: UniProt accession P04632 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 826 Human
      • Entrez Gene: 12336 Mouse
      • Entrez Gene: 29156 Rat
      • Omim: 114170 Human
      • SwissProt: P04632 Human
      • SwissProt: O88456 Mouse
      • SwissProt: Q64537 Rat
      • Unigene: 515371 Human
      • Unigene: 439782 Mouse
      • Unigene: 3430 Rat
      see all
    • 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 S1 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

    Images

    • Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
      Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
      All lanes : Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : CAPNS1 (Calpain small subunit 1) knockout HAP1 whole cell lysate
      Lane 3 : HepG2 whole cell lysate
      Lane 4 : HeLa whole cell lysate

      Lysates/proteins at 40 µg per lane.

      Predicted band size: 28 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab92356 observed at 82 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab92356 was shown to specifically react with Calpain 1 in wild-type HAP1 cells as signal was lost in CAPNS1 (Calpain small subunit 1) knockout cells. Wild-type and CAPNS1 (Calpain small subunit 1) knockout samples were subjected to SDS-PAGE. ab92356 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
      Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
      All lanes : Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356) at 1/1000 dilution

      Lane 1 : T47-D cell lysate
      Lane 2 : 293T cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : Human spleen lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit antibody at 1/2000 dilution

      Predicted band size: 28 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)

      ab92356 at 1/100 dilution staining Calpain small subunit 1 in paraffin-embedded Human kidney

      Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

    • Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)
      Anti-Calpain small subunit 1 antibody [EPR3323] (ab92356)

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (2)

    Publishing research using ab92356? Please let us know so that we can cite the reference in this datasheet.

    ab92356 has been referenced in 2 publications.

    • Seymen CM  et al. Melatonin Modulates NMDA-Receptor 2B/Calpain-1/Caspase-12 Pathways in Rat Brain After Long Time Exposure to GSM Radiation. Turk Neurosurg N/A:N/A (2019). WB, IHC-P ; Rat . PubMed: 31608966
    • Kim DH  et al. Calpain 9 as a therapeutic target in TGFß-induced mesenchymal transition and fibrosis. Sci Transl Med 11:N/A (2019). PubMed: 31316008

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