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No .

E-EL-R0013c

Rat IL-2(Interleukin 2) ELISA Kit

Item No:

E-EL-R0013c

Chinese Name:

Alias:

IL2, TCGF, Lymphokine

Keywords:

IHC kits

Classification:

IHC kits

Retail Price

¥ 2400

Market Price

¥ 2400

This kit is used for the in vitro quantification of IL-2 in Rat serum, plasma or other biological fluids

Spec
- 48T
- 96T
- 96T*5

Quantity

Inventory surplus

2997

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Describe

Use

　　This kit is used for the in vitro quantification of IL-2 in Rat serum, plasma or other biological fluids

Detection principle

　　This kit adopts double antibody sandwich ELISA method. Anti-Rat IL-2 antibody is used to coat the ELISA plate. The rat IL-2 in the sample or standard will bind to the coating antibody during the experiment and the free components are washed away. Biotinylated anti-rat IL-2 antibody and horseradish peroxidase-labeled affin were added sequentially. The anti-rat IL-2 antibody binds to the rat IL-2 bound to the encapsulated antibody, and the biotin binds specifically to the affin to form an immune complex, and the free components are washed away. The chromogenic substrate (TMB) was added, and the TMB appeared blue catalyzed by horseradish peroxidase and turned yellow after the addition of termination solution. The concentration of IL-2 in the sample was calculated by plotting the standard curve, using an enzyme marker to measure the OD value at 450 nm, with a positive ratio between IL-2 concentration and OD450 value.

Reaction Type	Sandwich method
Specification	96T
Reaction time	3.5h
Reactivity	Rat
Detection method	Colormetric
Detection range	31.25—2000 pg/mL
Sensitivity	18.75 pg/mL
Sample volume	100μL
Sample type	Serum, plasma or other biological fluids

Specificity

　　Detects rat IL-2 in samples with no significant cross-reactivity with other related proteins.

Repeatability

　　Within the plates, the coefficients of variation between the plates were <10%.

Typical Data

　　The OD values of the standard curves may vary due to different experimental operating conditions (e.g. operator, pipetting technique, plate washing technique and stabilization conditions, etc.). The following data and curves are for reference only. Experimenters need to establish standard curves according to their own experiments.

(pg/mL)	O.D	Average	Corrected
2000	2.496 2.516	2.506	2.439
1000	1.66 1.666	1.663	1.596
500	0.961 0.945	0.953	0.886
250	0.454 0.484	0.469	0.402
125	0.275 0.255	0.265	0.198
62.5	0.176 0.16	0.168	0.101
31.25	0.111 0.127	0.119	0.052
0	0.062 0.070	0.067	--

Precision

　　Intra-plate precision: 20 times on 1 plate for low, medium and high concentration samples.
　　Intra-plate precision: 20 times on 3 plates for low, medium and high concentrations.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	109.40	281.10	992.70	106.20	286.20	1010.30
Standard deviation	5.80	14.10	40.70	6.80	17.20	53.50
C V (%)	5.30	5.02	4.10	6.40	6.01	5.30

Recovery

　　Recovery experiments were performed by adding known concentrations of target proteins to five different samples, and the recovery range and average recovery were obtained.

Sample Type	Range (%)	Average Recovery (%)
Serum (n=8)	91-105	99
EDTA plasma (n=8)	95-109	102
Cell culture media (n=8)	93-110	101

Linearity

　　Recovery experiments were performed by adding known concentrations of target proteins to 5 samples, and the range of recovery rates and average recovery rates were obtained. Five samples were diluted 2, 4, 8, and 16 times for recovery experiments, and the range of recovery rates and average recovery rates were obtained.

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	93-108	97-113	92-106
1:2	Average (%)	100	104	98
1:4	Range (%)	98-112	85-96	94-110
1:4	Average (%)	106	90	101
1:8	Range (%)	101-114	80-91	94-107
1:8	Average (%)	108	87	99
1:16	Range (%)	99-112	82-95	94-104
1:16	Average (%)	106	88	99

Kit composition and storage

　　Unopened kits can be stored at 2-8°C for one week; if the kit is used after one week, please unpack the kit and store each component separately according to the conditions in the table below.

Chinese name	English Name	规格	保存条件
ELISA enzyme standard plate (detachable)	Micro ELISA Plate(Dismountable)	8 holes×12 strips	-20℃, can be stored for 6 months
Lyophilized standards	Reference Standard	2 sticks
Concentrated biotinylated antibody(100×)	Concentrated Biotinylated Detection Ab	1 x 120 μL
Concentrated HRP enzyme conjugate(100×)	Concentrated HRP Conjugate	1 x 120 μL	-20℃(keep away from light), can be stored for 6 months
Standard & Sample Diluent	Reference Standard & Sample Diluent	1 bottle of 20 mL	2-8℃. Can be stored for 6 months
Biotinylated antibody diluent	Biotinylated Detection Ab Diluent	1 bottle 14 mL
Enzyme Conjugate Diluent	HRP Conjugate Diluent	1 bottle 14 mL
Concentrated Washing Solution(25×)	Concentrated Wash Buffer (25×)	1 bottle 30 mL
Substrate solution (TMB)	Substrate Reagent	1 bottle 10 mL	2-8℃(keep away from light)
Reaction termination solution	Stop Solution	1 bottle 10 mL	2-8℃
Plate lamination	Plate Sealer	5 sheets
Product manual	Product Description	1 copy
Quality inspection report	Certificate of Analysis	1 copy

　　 Note: All reagent bottles must be tightly capped to prevent evaporation and microbial contamination.
　　The volume of reagents is subject to the actual shipping instructions. The relevant reagents will be slightly more than the volume indicated on the label when dispensing, please measure rather than pour directly when using.

Items required for the test

1. enzyme calibrator (450nm wavelength filter)

2. High precision pipettes, EP tubes and disposable tips: 0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL

3. 37°C thermostat, double-distilled water or deionized water

4. absorbent paper

Operation steps

	1. Add 100 μL of standard working solution or sample to the corresponding plate wells and incubate at 37°C for 90 minutes
	2. Immediately after discarding the liquid in the plate, add 100 μL of biotinylated antibody working solution and incubate at 37°C for 60 minutes
	3. Discard the liquid in the plate and wash the plate 3 times
	4. Add 100μL of HRP enzyme conjugate working solution to each well, incubate at 37℃ for 30 minutes, discard the liquid in the plate, and wash the plate 5 times
	5. Add 90μL of substrate solution to each well and incubate at 37℃ for 15 minutes
	6. Add 50μL of termination solution to each well
	7. Read immediately at 450 nm and process the data