ELISA Kit for Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) - USEB932Mu

ELISA Kit for Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH)

Size: 96T

Catalogue Number: USEB932Mu-96

Citations, Manuals and MSDS Available upon request.

Target: Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH)

Alternative Names: G3PD; GAPD; Peptidyl-cysteine S-nitrosylase GAPDH

Target Species: Mouse

Detection range: 0.156-10ng/mL

Sensitivity: 0.056ng/mL

Uniprot: P16858

Gene ID: 14433

Featured Series Function: Detects protein (regular version)

Recommended/Predicted Sample Types: Tissue Homogenates, Cell Lysates, Cell Culture Supernates and other Biological Fluids

Assay Time: 3h

Method: Colormetric

Specificity: Reactive with Mouse GAPDH / Glyceraldehyde-3-Phosphate Dehydrogenase

Detection principle: Double-antibody Sandwich

Sample Size: 100uL

Assay Precision: Intra-Assay: CV<10%, Inter-Assay: CV<12%

Storage: 4°C for 1 month/ -20°C for long-term(One year within shelf life)

Shelf-life: 12 months

Specificity: This assay has high sensitivity and excellent specificity for detection of Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH).

No significant cross-reactivity or interference between Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) and analogues was observed.

Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) were tested on 3 different plates, 8 replicates in each plate.

CV(%) = SD/meanX100

Stability: The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.

To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Assay procedure summary: 1. Prepare all reagents, samples and standards;

2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;

3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;

4. Aspirate and wash 3 times;

5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;

6. Aspirate and wash 5 times;

7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;

8. Add 50µL Stop Solution. Read at 450nm immediately.

Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Research Area: Enzyme & Kinase; Metabolic pathway; Cardiovascular biology; Hepatology; Nutrition metabolism;

Research Use Only