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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] - BSA and Azide free (ab247549)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)
  • Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)
  • Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)

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 - BSA and Azide free
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Conjugation
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PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)

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Overview

  • Product name

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

    Rabbit monoclonal [EPR3323] to Calpain small subunit 1 - BSA and Azide free
  • Host species

    Rabbit
  • Specificity

    Detects Calpain small subunit 1.

  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • General notes

    Ab247549 is the carrier-free version of ab92356. 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.

    ab247549 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Affinity purified
  • 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] (ab92356)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
  • Positive Controls

    • A431 membrane extract lysate (ab30134)
  • Recombinant Protein

    • Recombinant Human Calpain small subunit 1 protein (ab180298)

Applications

Our Abpromise guarantee covers the use of ab247549 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Use of a HRP/AP polymerized secondary antibody is recommended.

 

WB Use at an assay dependent concentration. Predicted molecular weight: 28 kDa.
  • 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] - BSA and Azide free (ab247549)
      Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)
      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



      This data was developed using ab92356, the same antibody clone in a different buffer formulation.

      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] - BSA and Azide free (ab247549)
      Western blot - Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)
      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



      This data was developed using ab92356, the same antibody clone in a different buffer formulation.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)
      This data was developed using ab92356, the same antibody clone in a different buffer formulation.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] - BSA and Azide free (ab247549)
      Anti-Calpain small subunit 1 antibody [EPR3323] - BSA and Azide free (ab247549)

    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

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (0)

    Publishing research using ab247549? Please let us know so that we can cite the reference in this datasheet.

    ab247549 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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