ELISA Kit for Cytochrome b-245 Beta Polypeptide (CYBb) - USED308Hu

ELISA Kit for Cytochrome b-245 Beta Polypeptide (CYBb)

Size: 96T

Catalogue Number: USED308Hu-96

Citations, Manuals and MSDS Available upon request.

Target: Cytochrome b-245 Beta Polypeptide (CYBb)

Alternative Names: CGD; GP91PHOX; NOX2; Neutrophil cytochrome b91; Cytochrome b558 beta; Heme-binding membrane glycoprotein gp91phox; Superoxide-generating NADPH oxidase heavy chain

Target Species: Human

Detection range: 0.156-10ng/mL

Sensitivity: 0.056ng/mL

Uniprot: P04839

Gene ID: 1536

Featured Series Function: Detects protein (regular version)

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

Assay Time: 3h

Method: Colormetric

Specificity: Reactive with Human CYBb / Cytochrome b-245 Beta Polypeptide

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 Cytochrome b-245 Beta Polypeptide (CYBb).

No significant cross-reactivity or interference between Cytochrome b-245 Beta Polypeptide (CYBb) and analogues was observed.

Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cytochrome b-245 Beta Polypeptide (CYBb) were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cytochrome b-245 Beta Polypeptide (CYBb) 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 Cytochrome b-245 Beta Polypeptide (CYBb). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Cytochrome b-245 Beta Polypeptide (CYBb). 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 Cytochrome b-245 Beta Polypeptide (CYBb), 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 Cytochrome b-245 Beta Polypeptide (CYBb) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Research Area: Metabolic pathway;

Research Use Only