Goat Anti-Rat Hypoxia inducible factor 1α
大鼠 (Rat) 低氧诱导因子1α(HIF-1α) 试剂盒
Storage: 2 - 8 °C Package size: 96 determinations
PRINCIPLE OF THE METHOD
The HIF-1α kit is a solid phase phase sandwich enzyme linked immuno sorbent assay(ELISA). Samples , including standards of known HIF-1α concentrations and unknowns are pipetted into these wells. During the first incubation, the HIF-1α antigen and a biotinylated monoclonal antibody specific for HIF-1α are simultaneously incubated. After washing, the enzyme(streptavidin-peroxydase)is added. After incubation and washing to remove all the unbound enzyme, a substrate solution which is acting on the bound enzyme is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of HIF-1α present in the samples.
REAGENTS PROVIDED AND RECONSTTTUTION
REAGENTS(Store at 2-8℃)
1×96 WELLS
0.5×96 WELLS
RECONSTTTUTION
96/48-wells microtiter plates
1
0.5
Ready-to-use
Plastiv cover
2
1
Ready-to-use
Standard: 800pg/ml
1Vials (0.6ml)
0.5Vials (0.3ml)
See reagents preparation on page 3
Blank control
1Vials (1.0ml)
1Vials (0.5ml)
Ready-to-use
Standard Diluent
1Vials (4.0ml)
1Vials (2.0ml)
Ready-to-use
Biotinylated anti-HIF-1α
1Vials (6.0ml)
1Vials (3.0ml)
Ready-to-use
Streptavidin-HRP
1Vials (8.0ml)
1Vials (4.0ml)
Ready-to-use
Washing Buffer
1Vials (20ml)
1Vials (10ml)
50× concentrate
Substrate A
1Vials (6.0ml)
1Vials (3.0ml)
Ready-to-use
Substrate B
1Vials (6.0ml)
1Vials (3.0ml)
Ready-to-use
Stopping Solution
1Vials (6.0ml)
1Vials (3.0ml)
Ready-to-use
Sample Diluent
1Vials (12ml)
1Vials (6.0ml)
Ready-to-use
MATERIAL REQUIRED BUT NOT PROVIDED
SAFETY
PROCEDURAL NOTES/LAB.QUALITY CONTROL
SPECIMEN COLLECTION\ PROCESSING AND STORAGE
PREPARATION OF REAGENTS
800 pg/ml
(6 Standard)
Original density 50ul。
400 pg/ml
(5 Standard)
100ul 6 Standard +100ul diludent
200 pg/ml
(4 Standard)
100ul 5 Standard +100ul diludent
100 pg/ml
(3 Standard)
100ul 4 Standard +100ul diludent
50 pg/ml
(2 Standard)
100ul 3 Standard +100ul diludent
25 pg/ml
(1 Standard)
100ul 2 Standard +100ul diludent
0 pg/ml
Blank Control
50ul。
ASSAY METHOD
SUGGESTED PLATE SCHEME
Standard
concentrations(pg/ml)
A
800
800
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
B
400
400
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
C
200
200
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
D
100
100
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
E
50
50
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
F
25
25
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
G
0
0
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
H
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
sample
LIMITATIONS OF THE PROCEDURE
Do not extrapolate the standard curve beyond the max standard curve point. The dose-response is non-linear in this region and good accruacy is difficult to obtain.
CALCULATION OF RESULTS
The minimum detectable concentration in this assay is estimated to be 1.0pg/ml