Rat Nppc (C-type natriuretic peptide) ELISA Kit

Mfr.No.

ER2085

Sensitivity

9.375pg/ml

Detection Range

15.6-1000pg/ml

Reactivity

Rat

Assay Duration

2 hours

Assay Type

Competitive

Description

This kit is based on Competitive-ELISA detection method and takes 2h assay time. The microplate provided in this kit has been precoated with Nppc. Nppc in the sample or standard competes with a fixed amount of biotin-labeled Nppc for sites on a pre-coated antibody specific to Nppc. Free components are washed off. After adding HRP-Streptavidin Conjugate (SABC), biotin specifically binds with streptavidin to form immune complexes. Wash off unbound conjugate. Add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that tued yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Compare OD450 value of sample with the standard curve through curve fitting software to determine the concentration of Nppc in the sample. The concentration of the target substance is inversely proportional to the OD450 value.

Alternative Names

C-type natriuretic peptide ELISA Kit, NPPC ELISA Kit, CNP2 ELISA Kit

Detection Method

Coated with Antigen

Kit Components

Kit Components	Item	Size(48T)	Size(96T)	Storage Condition for Opened Kit
E001	ELISA Microplate(Dismountable)	8x6	8x12	Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002	Lyophilized Standard	1vial	2vial	Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003	Biotin-labeled Antibody(Concentrated, 100X)	30ul	60ul	2-8°C (Avoid Direct Light)
E034	HRP-Streptavidin Conjugate(SABC, 100X)	60ul	120ul
E024	TMB Substrate	5ml	10ml
E039	Sample Dilution Buffer	10ml	20ml	2-8°C
E040	Antibody Dilution Buffer	5ml	10ml
E049	SABC Dilution Buffer	5ml	10ml
E026	Stop Solution	5ml	10ml
E038	Wash Buffer(Concentrated, 25X)	15ml	30ml
E006	Plate Sealer	3 pieces	5 pieces
E007	Product Description	1 copy	1 copy

Recovery

Add a certain amount of Nppc into the sample. Calculate the recovery by comparing the measured value with the expected amount of Nppc in the sample.

Sample Type	Recovery Range(%)	Average(%)
serum(n=10)	87-103	97
EDTA plasma(n=10)	88-100	95
Heparin plasma(n=10)	92-105	99

Linearity

Dilute the sample with a certain amount of Nppc at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type	1:2	1:4	1:8
serum(n=10)	85-104%	86-100%	87-100%
EDTA plasma(n=10)	89-101%	86-99%	87-100%
Heparin plasma(n=10)	85-101%	88-96%	86-97%

Specificity

Specifically binds with Nppc , no obvious cross reaction with other analogues.

Standard Curve

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.) Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(pg/ml)	OD-1	OD-2	Average
0	2.303	2.369	2.336
15.625	1.909	1.965	1.937
31.25	1.421	1.463	1.442
62.5	0.941	0.969	0.955
125	0.641	0.659	0.65
250	0.421	0.433	0.427
500	0.291	0.299	0.295
1000	0.173	0.179	0.176

Sample Type

Tissue lysate, Cell lysates, Cell culture supeatants, Plasma, Other biological fluids

Assay Procedure Summary

Step 1:
Wash the plate twice before adding the standard and sample.

Step 2:
Add 50ul standard or sample into each well. Immediately add 50ul biotin-labeled antibody working solution into each well(The tip can't be reused after touching the liquid.), gently shake the plate for 1min, seal closely and statically incubate for 45 minutes at 37°C.

Washing:
Wash the plate three times and immerse for 1min each time.

Step 3:
Add 100ul HRP-Streptavidin Conjugate (SABC) working solution into each well, seal the plate and statically incubate for 30 minutes at 37°C.

Washing:
Wash the plate five times and immerse for 1min each time.

Step 4:
Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)

Step 5:
Add 50ul stop solution. Read at 450nm immediately and calculate.

Precision

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/ml)	32.15	121.9	526.1	32.29	125.4	516
Standard deviation	1.48	6.14	28.09	1.53	6.56	26.26
CV(%)	4.59	5.04	5.34	4.73	5.23	5.09

Stability

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5)	37°C for 1 month	2-8°C for 6 months
Average(%)	80	95-100

Notes

Specifically binds with Nppc , no obvious cross reaction with other analogues.

Usage Notes

Step 1:
Take out the required plate wells, add 50ul Cap/Det Ab into each well, then add 50ul Standard or Sample into individual well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure thorough mixing then static incubate for 60 minutes at 37°C.

Washing:
Wash the plate twice without immersion.

Step 2:
Add 100ul HRP-Streptavidin (orange) into each well, seal the plate and static incubate for 30 minutes at 37°C.

Washing:
Wash the plate five times without immersion.

Step 3:
Add 90ul TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)

Step 4:
Add 50ul stop solution. Read at 450nm immediately and calculate.

UniProt ID	P55207

Storage	2-8°C(Sealed), Don't cryopreserve.

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SKU	Supplier No.	Size	Your price	Quantity
BHE10809740	ER2085	48T	$280		Add to Cart
BHE10809740	ER2085	96T	$400		Add to Cart

Rat Nppc (C-type natriuretic peptide) ELISA Kit

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