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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) (C terminal)
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
Alternative versions available:
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.
Our Abpromise guarantee covers the use of ab45689 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 17 kDa).|
|ICC/IF||1/100 - 1/250.|
|Flow Cyt||1/50 - 1/100.
ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IP||1/20 - 1/50.|
|IHC-Fr||Use at an assay dependent concentration.|
ab45689 immunoprecipitating Calmodulin. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP secondary antibody (HRP) (ab131366) at a dilution of 1/10000.
Lane 1: Human skeletal muscle lysate (10ug)
Lane 2: Human skeletal muscle lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab45689 in Human skeletal muscle lysate
ab45689 staining Calmodulin in NIH/3T3 (mouse embryo) 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)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
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 1: PBS in place of primary antibody.
Blocking and diluting buffer: 5% NFDM/TBST
Blocking and Diluting buffer: 5% NFDM/TBST
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. A Alexa Fluor® 555-conjugated Goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.
ICC/IF image of ab45689 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise 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.