Recombinant
RabMAb

Recombinant Anti-Calmodulin 1/2/3 antibody [EP799Y] - BSA and Azide free (ab214793)

Overview

  • Product name

    Anti-Calmodulin 1/2/3 antibody [EP799Y] - BSA and Azide free
    See all Calmodulin 1/2/3 primary antibodies
  • Description

    Rabbit monoclonal [EP799Y] to Calmodulin 1/2/3 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, ICC/IF, Flow Cyt, IHC-P, IHC-Fr, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    corresponding to Human Calmodulin 1/2/3 aa 100-200 (C terminal).

  • Positive control

    • NIH 3T3 cell lysate and human urinary bladder carcinoma.
  • General notes

    Ab214793 is the carrier-free version of ab45689. 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.

    ab214793 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab214793 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
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 16 kDa (predicted molecular weight: 17 kDa).

Target

  • Relevance

    Function: Calmodulin mediates the control of a large number of enzymes and other proteins by Ca(2+). Among the enzymes to be stimulated by the calmodulin-Ca(2+) complex are a number of protein kinases and phosphatases. Together with CEP110 and centrin, is involved in a genetic pathway that regulates the centrosome cycle and progression through cytokinesis.
  • Cellular localization

    Cytoplasm > cytoskeleton > spindle. Cytoplasm > cytoskeleton > spindle pole. Distributed throughout the cell during interphase, but during mitosis becomes dramatically localized to the spindle poles and the spindle microtubules.
  • Database links

  • Form

    There are three genes which encode an identical calcium binding protein which is one of the four subunits of phosphorylase kinase.
  • Alternative names

    • CALM 1 antibody
    • CALM 2 antibody
    • CALM 3 antibody
    • CALM antibody
    • CALM1 antibody
    • CALM2 antibody
    • CALM3 antibody
    • CALML2 antibody
    • calmodulin 1 (phosphorylase kinase, delta) antibody
    • Calmodulin 1 antibody
    • Calmodulin 2 (phosphorylase kinase, delta) antibody
    • Calmodulin 2 antibody
    • Calmodulin 3 (phosphorylase kinase, delta) antibody
    • Calmodulin 3 antibody
    • CAM 2 antibody
    • CAM 3 antibody
    • CAM antibody
    • CAM I antibody
    • CAM1 antibody
    • CAM2 antibody
    • CAM3 antibody
    • CAMB antibody
    • CAMC antibody
    • CAMI antibody
    • CAMII antibody
    • CAMIII antibody
    • CPVT4 antibody
    • DD132 antibody
    • FLJ99410 antibody
    • LP7057 protein antibody
    • PHKD antibody
    • PHKD2 antibody
    • PHKD3 antibody
    • phosphorylase kinase delta antibody
    • phosphorylase kinase, delta subunit antibody
    see all

Images

  • ab45689 immunoprecipitating Calmodulin. 10 µg of cell lysate was incubated with primary antibody at a dilution of 1/20. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

    Lane 1: Human skeletal muscle lysate (10 µg)
    Lane 2: Human skeletal muscle lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab45689 in human skeletal muscle lysate

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

  • ab45689 staining Calmodulin in NIH/3T3 (Mouse embryo fibroblast cell line) cells by flow cytometry.

    Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabeled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

  • ab45689 staining Calmodulin in human testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control: PBS in place of primary antibody (inset).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

  • ab45689 staining Calmodulin in mouse brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections).

    Tissue was fixed with acetone and blocked with 10% serum for 60 minutes at 4°C. Samples were incubated with primary antibody (1/100 in PBS + 0.1% Triton) for 12 hours at 4°C. An Alexa Fluor® 555-conjugated Goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

  • ab45689 at a 1:250 dilution staining Calmodulin in human urinary bladder carcinoma tissue by immunohistochemistry in paraffin embedded tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

  • ICC/IF image of ab45689 stained MCF7 (Human breast adenocarcinoma cell line) cells.

    The cells were fixed in 100% methanol for 5 minutes and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45689, 1/1000 dilution) overnight at +4°C.

    The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45689).

  • Overlay histogram showing MCF7 (Human breast adenocarcinoma cell line) cells stained with ab45689 (red line).

    The cells were fixed with 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 followed by the antibody (ab45689, 1/50 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 decreased signal in MCF7 cells fixed with 4% paraformaldehyde (10 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 (ab45689).

References

ab214793 has not yet been referenced specifically in any publications.

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