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GAPDH (Glyceraldehyde-3-phosphate Dehydrogenase) is an enzyme that is involved in the second phase of glycolysis, a process that breaks down glucose and ultimately produces pyruvate for the trichloroacetic acid cycle (TCA cycle), and adenosine triphosphate (ATP), which is the primary energy source for important biological functions.
GAPDH is generally thought to be present at a constant level within the cell. In most cases, this stays true regardless of experimental treatment or technical procedures. Because of its ability to remain present, the measurement of GAPDH is used as an internal control for experimental error. GAPDH is widely used in different assay methods to investigate certain aspects of cell activity.
The Human Total GAPDH ELISA Kit is used to measure the presence of human GAPDH in cell lysates.
Typical Data
This standard curve is provided for demonstration only as Fig.3. A standard curve should be generated for each set of samples assayed.
Sensitivity
The limit of detectable dose (LOD) of GAPDH is 0.15625ng/ml.
Specificity
This assay measures the total human/mouse/rat GAPDH in cell lysates.
For Research Use only
Simple Procedures (Fig. 1)
Simple Standard Preparation
Accurate Standard Curve
Kit Content and Storage Conditions
| Part | Part# | Description | Storage of Opened/Reconstituted |
|---|---|---|---|
| Human Total GAPDH Microplate | TBS32102A | 96 well microplate (12 strips of 8 wells) coated with a Capture Antibody specific for human GAPDH. | The unused wells can be stored in the sealed foil pouch containing the desiccant pack for up to 1 month at 2-8 °C. |
| Human GAPDH Standard | TBS32102B | 100 μL of Recombinant human GAPDH protein (200 ng/mL). | Aliquot and store at -20 ℃ for up to 1 month in a manual defrost the freezer. Avoid repeated freeze-thaw cycles. |
| Detection A | TBS32102C | 12 mL of human biotinylated GAPDH antibody. | May be stored for up to 3 months at 2-8 °C. |
| Detection B | TBS32102D | 12 mL of Streptavidin-HRP | |
| Assay Diluent | TBS32102E | 12 mL of a buffered protein base with preservatives. | |
| Wash Buffer | TBS3000W | 12 mL of concentrated solution (10x). | |
| TMB Substrate | TBS3000T | 12 mL of ultra-sensitive TMB substrate. | |
| Stop Solution | TBS3000S | 6 mL of 2 N sulfuric acid. |
Store the unopened kit at 2-8 ℃. Do not use past kit expiration dates.
The kit contains sufficient materials to run an ELISA on at least two 96-well plates.