Recombinant
RabMAb

Anti-Calpain 1 antibody [EPR3319] (ab108400)

Submit a review Q&A (1)References (6)

Overview

  • Product name
    Anti-Calpain 1 antibody [EPR3319]
    See all Calpain 1 primary antibodies
  • Description
    Rabbit monoclonal [EPR3319] to Calpain 1
  • Host species
    Rabbit
  • Specificity
    ab108400 recognises the large subunit of Calpain
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Calpain 1 aa 150-250. The exact sequence is proprietary.

  • Positive control
    • Human fetal kidney, U937, A431, (D) 293T, and T47D cell lysates; Human kidney tissue; HeLa cells.
  • 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 ab108400 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 1/1000 - 1/5000. Predicted molecular weight: 82 kDa.

For unpurified use at 1/1000 - 1/10000.
IHC-P 1/200.

See IHC antigen retrieval protocols.

For unpurified use at 1/500 - 1/1000.
Flow Cyt 1/380.

For unpurified use at 1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/50.

For unpurified use at 1/250 - 1/500.

Target

  • Function
    Calcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.
  • Tissue specificity
    Ubiquitous.
  • Sequence similarities
    Belongs to the peptidase C2 family.
    Contains 1 calpain catalytic domain.
    Contains 4 EF-hand domains.
  • Cellular localization
    Cytoplasm. Cell membrane. Translocates to the plasma membrane upon Ca(2+) binding.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • Ca2 activated neutral protease 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 1 antibody
    • Calpain 1 antibody
    • Calpain 1 large subunit antibody
    • Calpain 1, (mu/I) large subunit antibody
    • Calpain mu type antibody
    • Calpain mu-type antibody
    • Calpain regulatory subunit antibody
    • Calpain small subunit 1 antibody
    • Calpain, large polypeptide L1 antibody
    • Calpain-1 catalytic subunit antibody
    • Calpain-1 large subunit antibody
    • CAN1_HUMAN antibody
    • CANP 1 antibody
    • CANP antibody
    • CANP small subunit antibody
    • CANP1 antibody
    • CANPL 1 antibody
    • CANPL1 antibody
    • CAPN 1 antibody
    • CAPN1 antibody
    • Cell proliferation inducing protein 30 antibody
    • Cell proliferation-inducing gene 30 protein antibody
    • Micromolar Calpain antibody
    • Micromolar-calpain antibody
    • Mu Calpain antibody
    • muCANP antibody
    • muCL antibody
    see all

Images

  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    All lanes : Anti-Calpain 1 antibody [EPR3319] (ab108400)

    Lane 1 : Wild type HAP1 whole cell lysate
    Lane 2 : CAPN1 (Calpain 1) knockout HAP1 whole cell lysate
    Lane 3 : A431 whole cell lysate

    Predicted band size: 82 kDa



    Ab108400 was shown to recognize Calpain 1 in wild-type HAP1. A faint cross-reactive band was observed in  CAPN1 (Calpain 1) knockout cells at 82 kDa when over-exposed. Due to the immunogen’s homology with CAPN2 (Calpain 2 Uniprot: P17655), this band could correspond to the Calpain 2 protein. Please note that cross-reactivity with this protein has not been confirmed experimentally.

  • Flow Cytometry - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Flow Cytometry - Anti-Calpain 1 antibody [EPR3319] (ab108400)

    Overlay histogram showing Jurkat cells stained with ab108400 (red line) at 1/380 dilution. The cells were fixed with 2% paraformaldehyde. The secondary antibody used was a FITC conjugated goat anti-rabbit IgG at 1/150 dilution. Isotype control antibody (green line) was rabbit monoclonal IgG used under the same conditions.

  • Immunocytochemistry/ Immunofluorescence - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Immunocytochemistry/ Immunofluorescence - Anti-Calpain 1 antibody [EPR3319] (ab108400)

    ab108400 staining Calpain 1 in the T-47D cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/70). ab150077(1/500) an Alexa Fluor®488-conjugated Goat anti-rabbit IgG was used as the secondary antibody. Nuclei were counterstained with DAPI.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 1 antibody [EPR3319] (ab108400)

    ab108400 staining Calpain 1 in Human transitional cell carcinoma of bladder tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/200). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    All lanes : Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

    Predicted band size: 82 kDa

  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/1000 dilution + C6 Lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), HRP- conjugated at 1/1000 dilution

    Predicted band size: 82 kDa

  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/5000 dilution + T47-D Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), HRP- conjugated at 1/1000 dilution

    Predicted band size: 82 kDa

  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/5000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 82 kDa

  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    All lanes : Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/1000 dilution (unpurified)

    Lane 1 : Human fetal kidney cell lysate
    Lane 2 : U937 cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : 293T cell lysate
    Lane 5 : T47D cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 82 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 1 antibody [EPR3319] (ab108400)

    ab108400, unpurified, at 1/500 dilution staining Calpain in Human kidney by Immunohistochemistry, Paraffin-embedded tissue.

  • Immunocytochemistry/ Immunofluorescence - Anti-Calpain 1 antibody [EPR3319] (ab108400)
    Immunocytochemistry/ Immunofluorescence - Anti-Calpain 1 antibody [EPR3319] (ab108400)

    ab108400, unpurified, at 1/250 dilution staining Calpain in HeLa cells by Immunofluorescence.

References

This product has been referenced in:
  • Yu W  et al. Lens-specific conditional knockout of Msx2 in mice leads to ocular anterior segment dysgenesis via activation of a calcium signaling pathway. Lab Invest N/A:N/A (2019). Read more (PubMed: 30683901) »
  • Wang X  et al. A Label-Free Quantitative Proteomic Analysis of Mouse Neutrophil Extracellular Trap Formation Induced by Streptococcus suis or Phorbol Myristate Acetate (PMA). Front Immunol 9:2615 (2018). Read more (PubMed: 30555459) »
See all 6 Publications for this product

Publishing research using ab108400? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

Question

A customer have a problem with Calpain 1 antibody (ab108400) IHC
staining. Briefly, the antibody staining pattern is unexpected. You can
find the detailed protocol and staining photos in the attached files.

Read More
Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab108400.


How long did they perform the antigen retrieval in the pressure cooker? They may have over-retrieved so they can decrease the amount of time.


They should also omit any permeabilization agent like Triton X-100.


I see that the customer only blocked for 5 - 10 mins and this may be too short of a time. What buffer have they tried and at what percentage? They should block with 10% serum for 1 hr at RT.


They may also want to try diluting the antibody more to 1:2000 or 1:5000 and incubate that overnight at 4C instead of 1 hr at RT.


Have they run a no primary control?


Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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