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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-Calpain 1 抗体 [EPR3319] (ab108400)

Reviews (1)Q&A (1)References (20)
Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Flow Cytometry (Intracellular) - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Immunocytochemistry/ Immunofluorescence - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Immunocytochemistry/ Immunofluorescence - Anti-Calpain 1 antibody [EPR3319] (ab108400)
  • Anti-Calpain 1 antibody [EPR3319] (ab108400)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3319] to Calpain 1
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 750 Alkaline Phosphatase APC Carrier Free HRP PE

製品の概要

  • 製品名

    Anti-Calpain 1 antibody [EPR3319]
    Calpain 1 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR3319] to Calpain 1

  • 由来種

    Rabbit

  • 特異性

    This antibody recognises the large subunit of Calpain

  • アプリケーション

    適用あり: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • Human fetal kidney, U937, A431, (D) 293T, and T47D cell lysates; Human kidney tissue; HeLa cells.

  • 特記事項

    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.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab108400の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
1/1000 - 1/5000. Predicted molecular weight: 82 kDa.

For unpurified use at 1/1000 - 1/10000.
IHC-P
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/500 - 1/1000.
ICC/IF
1/50.

For unpurified use at 1/250 - 1/500.
Flow Cyt (Intra)
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.
特記事項
WB
1/1000 - 1/5000. Predicted molecular weight: 82 kDa.

For unpurified use at 1/1000 - 1/10000.
IHC-P
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/500 - 1/1000.
ICC/IF
1/50.

For unpurified use at 1/250 - 1/500.
Flow Cyt (Intra)
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.

ターゲット情報

  • 機能

    Calcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.

  • 組織特異性

    Ubiquitous.

  • 配列類似性

    Belongs to the peptidase C2 family.
    Contains 1 calpain catalytic domain.
    Contains 4 EF-hand domains.

  • 細胞内局在

    Cytoplasm. Cell membrane. Translocates to the plasma membrane upon Ca(2+) binding.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • 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

画像

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

    Overlay histogram showingJurkat 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.

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

参考文献 (20)

ab108400 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab108400 は 20 報の論文で使用されています。

  • Zhang S  et al. Calpain-2 Facilitates Autophagic/Lysosomal Defects and Apoptosis in ARPE-19 Cells and Rats Induced by Exosomes from RPE Cells under NaIO3 Stimulation. Oxid Med Cell Longev 2023:3310621 (2023). PubMed: 36703913
  • Liu L  et al. Dapagliflozin Ameliorates STZ-Induced Cardiac Hypertrophy in Type 2 Diabetic Rats by Inhibiting the Calpain-1 Expression and Nuclear Transfer of NF-κB. Comput Math Methods Med 2022:3293054 (2022). PubMed: 35096128
  • Kao SH  et al. Surface Viscosity-Dependent Neurite Initiation in Cortical Neurons. Adv Biol (Weinh) 6:e2101325 (2022). PubMed: 35362269
  • Wang P  et al. Pericardial Adipose Tissue-Derived Leptin Promotes Myocardial Apoptosis in High-Fat Diet-Induced Obese Rats Through Janus Kinase 2/Reactive Oxygen Species/Na+/K+-ATPase Signaling Pathway. J Am Heart Assoc 10:e021369 (2021). PubMed: 34482701
  • Wang X  et al. Piezo type mechanosensitive ion channel component 1 facilitates gastric cancer omentum metastasis. J Cell Mol Med 25:2238-2253 (2021). PubMed: 33439514
View all Publications for this product

レビューと Q&A

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1-2 of 2 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-Calpain 1 antibody [EPR3319]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Rabbit Tissue sections (Hippocampus (Neurons in the CA1 Region))
Permeabilization
Yes - 0,1% Triton in PBS
Specification
Hippocampus (Neurons in the CA1 Region)
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Helena Lichtenfeld

Verified customer

投稿 Aug 18 2020

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.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com