製品の概要

  • 製品名Anti-Calmodulin antibody [2D1]
    Calmodulin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [2D1] to Calmodulin
  • 特異性This antibody detects calmodulin. It does not detect parvalbumin, tropinin, S-100, or myosin light chain kinase (MLCK).By Western blot, this antibody detects a 17 kDa protein representing calmodulin from Dictyostelium cell lysate. Immunohistochemical staining of calmodulin in Dictyostelium cells with this antibody results in staining of the contractile vacuoles.
  • アプリケーション適用あり: Flow Cyt, ELISA, WB, IHC-P, ICC, IP, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Chicken, Cow, Human, Dictyostelium discoideum
    交差が予測される動物種: Wheat
  • 免疫原

    Other Immunogen Type corresponding to Calmodulin. Calmodulin purified from Dictyostelium discoideum.

  • ポジティブ・コントロール
    • WB: Purified Dictyostelium calmodulin IHC: rat brain
  • 特記事項

     

     

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab2860 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Flow Cyt Use 2µg for 106 cells.
ELISA Use at an assay dependent concentration.
WB 1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).
IHC-P 1/20.
ICC 1/50.
IP Use at an assay dependent concentration.
ICC/IF 1/20.

ターゲット情報

Anti-Calmodulin antibody [2D1] 画像

  • Flow cytometry analysis of Calmodulin showing positive staining in the cytoplasm of PC12 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2860 at 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
  • Flow cytometry analysis of Calmodulin showing positive staining in the cytoplasm of MCF-7 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2860 at a dilution of 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
  • Flow cytometry analysis of Calmodulin showing positive staining in the cytoplasm of C6 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2860 at 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
  • Immunolocalization of calmodulin in rat brain using ab2860 (1:100)
  • Anti-Calmodulin antibody [2D1] (ab2860) at 1/500 dilution + Purified Dictyostelium calmodulin

    Predicted band size : 17 kDa
  • Overlay histogram showing HeLa cells stained with ab2860 (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. The cells were then incubated with the antibody (ab2860, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunofluorescent analysis of Calmodulin using Calmodulin Monoclonal antibody (2D1) ab2860 shows staining in HeLa cells. Calmodulin staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Calmodulin ab2860 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of Calmodulin using Calmodulin Monoclonal antibody (2D1) ab2860 shows staining in A2058 melanoma cells. Calmodulin staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Calmodulin ab2860 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of Calmodulin using Calmodulin Monoclonal antibody (2D1) ab2860 shows staining in C6 glioma cells. Calmodulin staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Calmodulin ab2860 at a dilution of 1:20 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Rat testis tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Calmodulin ab2860 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Rat cerebellum tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Calmodulin ab2860 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Anti-Calmodulin antibody [2D1] (ab2860) 使用論文

This product has been referenced in:
  • Li RK  et al. Iqcg is essential for sperm flagellum formation in mice. PLoS One 9:e98053 (2014). ICC/IF ; Mouse . Read more (PubMed: 24849454) »
  • Boraldi F  et al. Hypoxia influences the cellular cross-talk of human dermal fibroblasts. A proteomic approach. Biochim Biophys Acta 1774:1402-13 (2007). WB ; Human . Read more (PubMed: 17904921) »

See all 3 Publications for this product

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Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

Unter unseren anti-Calmodulin- Antikörpern ist ab2860 vermutlich der aussichtsreichste Kandidat: Das Immunogen (von Dictyostelium discoideum, SwissProtID P...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Schizosaccharomyces pombe Recombinant protein (N/A)
Loading amount 10 µg
Specification N/A
Gel Running Conditions Reduced Denaturing (15%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
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Abcam user community

Verified customer

投稿 Mar 09 2011

Thank you for your phone enquiry and questionaire. I'm sorry to hear you are having a problem with ab2860 (calmodulin Ab) in Western blotting. We have received other enquiries regarding this antibody and I would like to suggest the following modifi...

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Thank you for your enquiry. I browsed the reference, Cell Mot. Cytoskel., 18: 113-122, 1991. There are a few suggestions, taken from the article, that I can give. These researchers had difficulty with their proteins transferring through the memb...

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I'm very sorry to hear you are experiencing problems with ab2860. I would also expect the lysates in rat and E.coli samples to work with this antibody, however there may be several reasons for the lack of signal you are currently experiencing. ...

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Thank you for those extra details about your protocol this helped me a lot as I looked deeper into the cross reactivity of this antibody with your samples: 293T and HeLa cells are from human origin and 3T3 cells are from mouse origin. This could be ...

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I'm sorry to hear you are having a problem with ab2860. We have not had a complaint about this antibody before. Thank you for taking the time to fill in our questionnaire, it helped me to understand your problem and it is useful to know that another...

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Thank you for your enquiry and for your patience. Regarding the specific epitope mapping, we have not performed these studies. Both immunogens were: Calmodulin purified from Dictyostelium discoideum and both antibodies have been tested for application ...

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