Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E89] to Caldesmon/CDM - BSA and Azide free
  • Suitable for: Flow Cyt, ICC/IF, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free
    See all Caldesmon/CDM primary antibodies

  • Description

    Rabbit monoclonal [E89] to Caldesmon/CDM - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Rat
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HeLa and NIH/3T3 cell lysates.

  • General notes

    ab215275 is the carrier-free version of ab32330 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

    Ab215275 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product was previously labelled as Caldesmon

    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

Images

  • Western blot - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Western blot - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    All lanes : Anti-Caldesmon/CDM antibody [E89] (ab32330) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : CALD1 knockout HeLa cell lysate
    Lane 3 : NIH/3T3 cell lysate
    Lane 4 : HEK-293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 93 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab32330).

    Lanes 1-4: Merged signal (red and green). Green - ab32330 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.

    ab32330 Anti-Caldesmon/CDM antibody [E89] was shown to specifically react with Caldesmon/CDM in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265026 (knockout cell lysate ab257375) was used. Wild-type and Caldesmon/CDM knockout samples were subjected to SDS-PAGE. ab32330 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Immunocytochemistry/ Immunofluorescence - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)

    Immunocytochemistry/Immunofluorescence analysis of C6 (rat glioma) cells labelling Caldesmon/CDM with purified ab32330 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei couterstained with DAPI (blue). 

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32330).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)

    ab32300, at a 1/250 dilution, staining human Caldesmon/CDM in uterus tissue by Immunohistochemistry, Paraffin embedded tissue

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32330).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Immunocytochemistry/ Immunofluorescence - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)

    ab32330, at a 1/250 dilution, staining human Caldesmon/CDM in HeLa cells by Immunofluorescence

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32330).

  • Flow Cytometry - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Flow Cytometry - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Overlay histogram showing HeLa cells stained with ab32330 (red line). The cells were fixed with 4% paraformaldehyde (10 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 followed by the antibody (ab32330, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HeLa cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32330).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)

    Fluorescent immunohistochemical analysis of paraffin-embedded human normal kidney vessels using ab32330. Green-Caldesmon/CDM red-PI

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32330).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)

    Fluorescent immunohistochemical analysis of paraffin-embedded human tissue using ab32330. Green-Caldesmon/CDM red-PI.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32330).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)

    Fluorescent immunohistochemical analysis of paraffin-embedded human normal uterus tissue using ab32330. Green-Caldesmon/CDM red-PI.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32330).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • OI-RD Scanning - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    OI-RD Scanning - Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32330).

  • Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)
    Anti-Caldesmon/CDM antibody [E89] - BSA and Azide free (ab215275)

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