ELISA Kit for Hemoglobin (HB)

Brand

Cloud-Clone

short description

Enzyme-linked immunosorbent assay for Antigen Detection.
Research focus: Metabolic pathway; Infection immunity; Hematology; Neuro science; ;

spezie

Cavia (Guinea pig )

Cross Reaction

This assay has high sensitivity and excellent specificity for detection of Hemoglobin (HB).
No significant cross-reactivity or interference between Hemoglobin (HB) and analogues was observed.

detection target

Hemoglobin

alias

Hgb; Haemoglobin; Heterotetramer(??)2

sensitivity

The minimum detectable dose of this kit is typically less than 4.58 ng/mL

detection range

12.50-800 ng/mL

certificate

ISO 9001:2008, ISO 13485:2003

sample type

Serum, plasma, erythrocyte lysates.

pack size

48T, 96T, 5x96T, 10x96T

Reproducibility

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hemoglobin (HB) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Hemoglobin (HB) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Assay time

3h

storage and Stability

All the kits should stored according to the manual.
The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 degree upon receipt while the others should be at 4 degree.
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.

Principle of the Elisa

Double-antibody Sandwich
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 Hemoglobin (HB). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Hemoglobin (HB). 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 Hemoglobin (HB), 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 Hemoglobin (HB) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Kit Content

Pre-coated, ready to use 96-well strip plate: 1
Plate sealer for 96 wells: 4
Standard: 2
Standard Diluent: 1x20mL
Detection Reagent A: 1x120µ L
Assay Diluent A: 1x12mL
Detection Reagent B: 1x120µ L
Assay Diluent B: 1x12mL
TMB Substrate: 1x9mL
Stop Solution: 1x6mL
Wash Buffer (30 x concentrate): 1x20mL
Instruction manual: 1

Assay procrdure summery

1. Prepare all reagents, samples and standards;
2. Add 100µ L standard or sample to each well. Incubate 2 hours at 37° C;
3. Aspirate and add 100µ L prepared Detection Reagent A. Incubate 1 hour at 37° C;
4. Aspirate and wash 3 times;
5. Add 100µ L prepared Detection Reagent B. Incubate 30 minutes at 37° C;
6. Aspirate and wash 5 times;
7. Add 90µ L Substrate Solution. Incubate 10-20 minutes at 37° C;
8. Add 50µ L Stop Solution. Read at 450nm immediately.