Mouse CitH3 (Histone H3 Citrullination) ELISA Kit

Mfr.No.

EM2158

Description

This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti CitH3 antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with CitH3 bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of CitH3 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Alternative Names

CitH3 ELISA Kit, Histone H3 Citrullination ELISA Kit

Reactivity

Mouse

Detection Method

Double Antibody

Detection Range

31.25-2000pg/ml

Sample Type

Cell lysates, Tissue lysate, Other liquid samples, Serum, Plasma, Cell culture supernatants

Assay Type

Sandwich

Assay Duration

4 hours

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)	60ul	120ul	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 CitH3 into the sample. Calculate the recovery by comparing the measured value with the expected amount of CitH3 in the sample.

Sample Type	Recovery Range(%)	Average(%)
serum(n=10)	87-97	94
EDTA plasma(n=10)	88-102	96
Heparin plasma(n=10)	87-104	98

Linearity

Dilute the sample with a certain amount of CitH3 at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type	1:2	1:4	1:8
serum(n=10)	91-99%	85-98%	87-104%
EDTA plasma(n=10)	83-96%	82-98%	93-98%
Heparin plasma(n=10)	80-92%	80-99%	80-92%

Sensitivity

18.75pg/ml

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	0.092	0.094	0.093
31.25	0.188	0.194	0.191
62.5	0.263	0.271	0.267
125	0.476	0.49	0.483
250	0.623	0.641	0.632
500	1.071	1.103	1.087
1000	1.599	1.645	1.622
2000	2.156	2.218	2.187

Assay Procedure Summary

Step 1:
Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.

Washing:
Wash the plate twice without immersing.

Step 2:
Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 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, 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.

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 CitH3 , no obvious cross reaction with other analogues.

Usage Notes	Microplate reader (wavelength: 450nm) 37°C incubator (CO2 incubator for cell culture is not recommenced.) Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose) Precision single (0.5-10uL, 5-50uL, 20-200uL, 200-1000uL) and multi-channel pipette with disposable tips(Calibration is required before use.) Sterile tubes and Eppendorf tubes with disposable tips Absorbent paper and loading slot Deionized or distilled water

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)	64.29	253.3	1005.8	62.73	242.2	1021.9
Standard deviation	3.26	13.3	53.31	3.27	12.28	53.45
CV(%)	5.07	5.25	5.3	5.21	5.07	5.23

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

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SKU	Supplier No.	Size	Your price	Quantity
BHE10809973	EM2158	48T	$280		Add to Cart
BHE10809973	EM2158	96T	$400		Add to Cart

Mouse CitH3 (Histone H3 Citrullination) ELISA Kit

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