- Predicted Species Reactivity:
- Kit Detection Method:
- Colorimetric, OD450 nm
- Kit Duration:
- ~ 3 Hours
- Kit Principle:
- Aviva Systems Biology CKB ELISA Kit (Mouse) (OKCD07926) is based on standard sandwich enzyme-linked immuno-sorbent assay technology. An antibody specific for CKB has been pre-coated onto a 96-well plate (12 x 8 Well Strips) and blocked. Standards or test samples are added to the wells, incubated and removed. A biotinylated detector antibody specific for CKB is added, incubated and followed by washing. Avidin-Peroxidase Conjugate is then added, incubated and unbound conjugate is washed away. An enzymatic reaction is produced through the addition of TMB substrate which is catalyzed by HRP generating a blue color product that changes to yellow after adding acidic stop solution. The density of yellow coloration read by absorbance at 450 nm is quantitatively proportional to the amount of sample CKB captured in the well.
- Kit Range:
- Kit Reproducibility:
- Intra-assay reproducibility was evaluated using 20 replicates of 3 samples representing low, middle and high level CKB. Inter-assay reproducibility was evaluated with 3 samples representing low, middle and high level CKB using 8 replicates on each of 3 plates.
Intra-Assay CV <10%
Inter-Assay CV <12%
- Kit Component:
Component Amount CKB Microplate 96 Wells (12 x 8 Well strips) CKB Lyophilized Standard 2 100X CKB HRP-Detector Anitbody 1 x 120 µL 100X Avidin-HRP Conjugate 1 x 120 µL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 2 mL Stop Solution 1 x 20 mL
- Reconstitution and Storage:
- Store as indicated in product manual.
- Gene Symbol:
- Official Gene Full Name:
- creatine kinase B
- Alias Symbols:
- Bck, Ck3, B-CK, Ck-3, Ckbb, CPK-B
- NCBI Gene Id:
- Protein Name:
- creatine kinase B-type
- Description of Target:
- Reversibly catalyzes the transfer of phosphate between ATP and various phosphogens (e.g. creatine phosphate). Creatine kinase isoenzymes play a central role in energy transduction in tissues with large, fluctuating energy demands, such as skeletal muscle, heart, brain and spermatozoa.
- Swissprot Id:
- Protein Accession #:
- Nucleotide Accession #:
- Sample Type:
- Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- < 30pg/mL
- Tissue Tool:
- Find tissues and cell lines supported by RNA-seq analysis to express CKB.
- RNA Seq:
- Find tissues and cell lines supported by DNA array analysis to express CKB.
- Click here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
- Tips Information:
Washington University in St. Louis
1. What sample types were used with this kit?
The samples used in this kit are young mouse lenses from different genotypes.
2. Were these samples validated or characterized as positive and/or negative controls?
We did not use any additional controls. Only those provided by the kit.
3. What dilution worked for your samples?
Samples were not diluted but each lens is only a 1-2 mg and they were homogenized in 0.5 mL 1X PBS.
4. Please provide a brief summary of your sample preparation method:
Retrieve Mice and dissect lenses. Rinse lenses in 1X PBS and homogenize on ice with 0.5 mL of 1X PBS. Store overnight at -20°C. Perform 2 freeze-thaw cycles. Centrifuge the homogenate for 20 min at 10,000 rpm. Assay immediately.