Bovine Histamine ELISA Kit
CELI-66051b 96 Tests
Operating instructions
FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS!PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE
BEGINNING!
Synonyms
Histamine, HIS
Search name
Bovine Histamine ELISA Kit, Bovine HIS ELISA Kit, Histamine ELISA Kit, HIS ELISA Kit
Intended use
Bovine Histamine ELISA Kit allows for the in vitro quantitative determination of general histamine,Histamine concentrations in serum, plasma,
tissue homogenates, cell culture supernates, and other biological fluids.
Test principle
Bovine Histamine ELISA Kit is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been
pre-coated with an antibody specific to Histamine, During the reaction, Histamine in the sample or standard competes with a fixed amount
of biotin-labeled Histamine for sites on a pre-coated Monoclonal antibody specific to Histamine. Excess conjugate and unbound sample or
standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and
incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a
sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of
Histamine in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Materials and components
Reagent Quantity
Assay plate 1
Standard 2
Sample Diluent 1 × 20ml
Assay Diluent A 1 × 10ml
Assay Diluent B 1 × 10ml
Detection Reagent A 1 × 60μl
Detection Reagent B 1 × 120μl
Wash Buffer(25 x concentrate) 1 × 30ml
Substrate 1 × 10ml
Stop Solution 1 × 10ml
Plate sealer for 96 wells 5
instructions 1
Other supplies required
Microplate reader.
Pipettes and pipette tips.
EP tube
Deionized or distilled water.
Storage of the kits
The Assay Plate, Standard, Detection Reagent A and Detection Reagent B should be stored at -20℃ upon being received. After receiving the kit , Substrate should be always stored at 4℃.Other reagents are kept according to the labels on vials. But for long term storage, please keep the whole kit at -20℃(Except the substrate). The unused strips should be kept in a sealed bag with the desiccant provided to minimize exposure to damp air. The test kit may be used throughout the expiration date of the kit (six months from the date of manufacture). Opened test kits will remain stable until the expiring date shown, provided it is stored as prescribed above.
Note: Because of the inherent stability of lyophilized material, EIAab may ship these materials at ambient temperature.
Performance Characteristics
Intra-Assay CV: ≤4.2
Inter-Assay CV: ≤7.3
Spike Recovery: 106%
Quick Reference Procedure
IMPORTANT: Please refer to the supplied protocol for preparation of Working Stocks as noted below.
General Histamine Standard Prepaparation(step 2)
Tube
|
Working Stock
|
S1
|
S2
|
S3
|
S4
|
S5
|
S6
|
S7
|
BL
|
Amount Taken from Tube to Left(ml)
|
---
|
0
|
0.5
|
0.5
|
0.5
|
0.5
|
0.5
|
0.5
|
---
|
Sample Diluent
|
---
|
1.0
|
0.5
|
0.5
|
0.5
|
0.5
|
0.5
|
0.5
|
1.0
|
Final Conc.(ng/ml)
|
500
|
500
|
250
|
125
|
62.5
|
31.2
|
15.6
|
7.8
|
0
|
Antibody Concentrate Preparation (Step 5)
Microplate Strips Used
|
2
|
4
|
6
|
8
|
10
|
12
|
Working Stock(ul)
|
20
|
40
|
60
|
80
|
100
|
120
|
Conjugate Diluent(ml)
|
2.0
|
4.0
|
6.0
|
8.0
|
10.0
|
12.0
|
HRP Conjugate Concentrate (step 7)
Microplate Strips Used
|
2
|
4
|
6
|
8
|
10
|
12
|
Working Stock(ul)
|
20
|
40
|
60
|
80
|
100
|
120
|
Conjugate Diluent(ml)
|
2.0
|
4.0
|
6.0
|
8.0
|
10.0
|
12.0
|
Caution:
Product is for research use only!