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No .

E-EL-M0828c

Mouse NGAL(Neutrophil Gelatinase Associated Lipocalin) ELISA Kit

Item No:

E-EL-M0828c

Chinese Name:

Alias:

LCN2, Lipocalin 2, Oncogene 24p3, MSFI

Keywords:

Reagent Kits

Classification:

Reagent Kits

Retail Price

¥ 3200

Market Price

¥ 3200

This kit is designed for the in vitro quantitative analysis of NGAL in Mouse serum, plasma or other biological fluids.

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

Quantity

Inventory surplus

2997

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Describe

Use

　　This kit is designed for the in vitro quantitative analysis of NGAL in Mouse serum, plasma or other biological fluids.

Detection Principle

　　This kit uses double antibody sandwich ELISA method. Anti-Mouse NGAL antibody is used to encapsulate on an enzyme labeling plate. Mouse NGAL in the sample or standard will bind to the encapsulated antibody during the experiment, and the free components will be washed away. Biotinylated anti-mouse NGAL antibody and horseradish peroxidase-labeled affinity protein are added sequentially. The anti-mouse NGAL antibody binds to mouse NGAL bound to the coated antibody, the biotin binds specifically to the affinity element to form an immune complex, and the free components are washed away. Colorimetric substrate (TMB) is added, and TMB shows blue color catalyzed by horseradish peroxidase, and turns into yellow color after addition of termination solution. The OD value was measured at 450 nm with an enzyme meter, and there was a positive relationship between the NGAL concentration and the OD450 value, and the concentration of NGAL in the sample was calculated by plotting the standard curve.

Reaction type	夹心法
Specification	96T
Reaction time	3.5h
Reactivity	Mouse
Detection method	Colormetric
Detection range	39.06—2500 pg/mL
Sensitivity	23.44 pg/mL
Sample Volume	100μL
Sample Type	血清，血浆或其他生物体液

Idiosyncrasy

　　Detects mouse NGAL in samples with no significant cross-reactivity with other related proteins.

Repeatable

　　The coefficients of variation within plates and between plates were all <10%.

Typical data

　　The OD values of the standard curve will 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, and the experimenter needs to establish the standard curve according to his/her own experiment.

(pg/mL)	O.D	Average	Corrected
2500	2.17 2.218	2.194	2.136
1250	1.493 1.513	1.503	1.445
625	0.768 0.756	0.762	0.704
312.5	0.431 0.453	0.442	0.384
156.25	0.229 0.221	0.225	0.167
78.13	0.146 0.144	0.145	0.085
39.06	0.094 0.114	0.104	0.046
0	0.054 0.062	0.058	--

Precision

　　 Intra-plate precision: Low, medium and high concentration samples were tested 20 times on 1 plate.
　　Inter-plate precision: low concentration, medium concentration and high concentration samples were tested 20 times on 3 plates.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	119.40	390.07	1093.66	121.64	352.20	1131.18
Standard deviation	6.93	17.28	43.64	7.99	18.81	60.41
C V (%)	5.80	4.43	3.99	6.57	5.34	5.34

Recovery rate

　　A known concentration of the target protein was added to five different samples and recovery experiments were done to obtain a range of recoveries and an average recovery.

Sample Type	Range (%)	Average Recovery (%)
Serum (n=8)	91-105	97
EDTA plasma (n=8)	89-102	95
Cell culture media (n=8)	91-108	99

linearly

　　Five samples were spiked with a known concentration of the target protein and recovery experiments were performed to determine the range of recovery and the average recovery. The 5 samples were diluted 2-fold, 4-fold, 8-fold, and 16-fold to obtain the recovery range and average recovery.

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	90-106	94-107	89-104
1:2	Average (%)	97	99	96
1:4	Range (%)	90-104	86-96	83-95
1:4	Average (%)	97	91	89
1:8	Range (%)	89-102	79-93	89-104
1:8	Average (%)	94	86	95
1:16	Range (%)	87-99	88-99	86-99
1:16	Average (%)	92	93	91

Kit Composition and Storage

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

Chinese name	English name	Specification	Storage conditions
ELISA酶标板(可拆卸)	Micro ELISA Plate(Dismountable)	8 holes×12 strips	-20℃, can be stored for 6 months
冻干标准品	Reference Standard	2 sticks
浓缩生物素化抗体(100×)	Concentrated Biotinylated Detection Ab	1 x 120 μL
浓缩HRP酶结合物(100×)	Concentrated HRP Conjugate	1 x 120 μL	-20℃ (avoid light), can be stored for 6 months
标准品&样品稀释液	Reference Standard & Sample Diluent	1 bottle 20 mL	2-8℃. Can be stored for 6 months
生物素化抗体稀释液	Biotinylated Detection Ab Diluent	1 bottle 14 mL
酶结合物稀释液	HRP Conjugate Diluent	1 bottle 14 mL
浓缩洗涤液(25×)	Concentrated Wash Buffer (25×)	1 bottle 30 mL
底物溶液(TMB)	Substrate Reagent	1 bottle 10 mL	2-8℃(avoid light)
反应终止液	Stop Solution	1 bottle 10 mL	2-8℃
封板覆膜	Plate Sealer	5 sheets
产品说明书	Product Description	1 sheet
质检报告	Certificate of Analysis	1 sheet

　　Note: All reagent bottle caps must be screwed on tightly to prevent evaporation and microbial contamination.
　　Reagent volumes are based on the actual shipping version of the instructions. Reagents may be dispensed in a slightly larger volume than indicated on the label, so please measure rather than pour when using.

Self-contained items required for testing

•1. Enzyme Labeling Instrument (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℃ thermostat, double-distilled water or deionized water

•4. absorbent paper

Procedure

	1. Add 100μL of standard working solution or sample into the corresponding plate wells and incubate at 37℃ for 90 minutes.
	2. Discard the liquid in the plate, immediately add 100μL of biotinylated antibody working solution and incubate at 37℃ 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 five 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. Immediately read at 450nm and process the data.