Assay total GSK-3β (pan glycogen synthase kinase 3 beta, GSK-3b + pGSK-3b) levels in cell lysates of human and rat origin with our Thermo Scientific Total GSK-3 beta ELISA Kit.
This GSK-3 beta ELISA kit is an immunoassay for quantitative determination of glycogen synthase kinase 3 beta (pan or total GSK-3β) in cell lysates. A recombinant GSK-3β standard is provided in the kit. The kit uses a monoclonal antibody to GSK-3β precoated in a microplate to bind the GSK-3β in the standards or sample. After a short incubation, the excess sample or standard is washed out and a rabbit polyclonal antibody to GSK-3β is added. This antibody binds to GSK-3β captured on the plate. The polyclonal antibody detects a non-phosphorylated region of GSK-3β and therefore detects GSK-3β regardless of phosphorylation status. After a short incubation, the excess antibody is washed out and secondary conjugate is added, which binds to the polyclonal GSK-3β antibody. Excess conjugate is washed out and substrate is added. After a short incubation, the enzyme reaction is stopped and the color generated is read at 450nm. The measured optical density is directly proportional to the concentration of GSK-3β in either standards or samples.
Highlights:
- Target: phosphorylated GSK-3β ([pSer9] glycogen synthase kinase 3 beta, pGSK-3β)
- Format: colorimetric ELISA (TMB substrate) in clear 96-well strip plate
- Measurement: absorbance at 450nm minus absorbance at 550nm
- Assay Range: 78.1 to 5000pg/mL
- Sensitivity: < 74.4pg/mL
- Precision: dependent upon sample type and level (see instructions)
- Sample Types: cell lysates of human, mouse, and rat origin
- Total Assay Time: 3 hours
- Sample Size: 100μL
- Specificity: no significant cross-reaction with GSK-3α, ATP Citrate Lyase, MEK-1, AKT, phospho-JNK, ERK and phospho-ERK.
Includes:
Complete kit contains pre-coated antibody-plate, detection antibodies, buffers, standard, substrate, and stop solution.
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