Human CitH3 (Histone H3 Citrullination) QuickTest ELISA Kit

Catalogue No.: QT-EH5022

Alias: CitH3 ELISA Kit, Histone H3 Citrullination ELISA Kit

Species: Human

Range: 31.25-2000pg/ml

Sensitivity: 18.75pg/ml

 

Product NameHuman CitH3 (Histone H3 Citrullination) QuickTest ELISA KitAliasCitH3 ELISA Kit, Histone H3 Citrullination ELISA KitCatalogue No.QT-EH5022Size48T/96TSpeciesHumanSample TypeSerum, plasma, cell culture supernatant, cell lysate or tissue lysate, other biological fluid samplesDetection MethodSandwich ELISA, Double AntibodyDetection WavelengthOD450Reaction Duration120 minutesRange31.25-2000pg/mlSensitivity18.75pg/mlStorage2-8°C(Sealed), Don't cryopreserve.SpecificitySpecifically binds with CitH3 , no obvious cross reaction with other analogues.ELISA Kit Components

Kit Components	Item	Size(48T)	Size(96T)	Storage Condition for Opened Kit
E001	ELISA Microplate(Dismountable)	8×6	8×12	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
E054	Cap/Det Ab (Ready to use, blue)	3ml	6ml	2-8°C (Avoid Direct Light)
E053	HRP-Streptavidin (Ready to use, orange)	5ml	10ml
E024	TMB Substrate	5ml	10ml
E039	Sample Dilution Buffer	20ml	20ml	2-8°C
E026	Stop Solution	5ml	5ml
E038	Wash Buffer(25X)	15ml	30ml
E006	Plate Sealer	3 pieces	5 pieces
E007	Product Description	1 copy	1 copy

Required Instruments and Reagents

1. Microplate reader (wavelength: 450nm)
2. 37°C incubator (CO2 incubator for cell culture is not recommenced.)
3. Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
4. Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
5. Sterile tubes and Eppendorf tubes with disposable tips
6. Absorbent paper and loading slot
7. Deionized or distilled water

Assay Procedure Summary

• 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.

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.

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)	87-105	98
EDTA plasma(n=10)	88-99	95
Heparin plasma(n=10)	88-104	97

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)	87-102%	88-95%	88-99%
EDTA plasma(n=10)	93-105%	90-100%	90-94%
Heparin plasma(n=10)	86-99%	92-97%	91-97%

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)	64.32	253.6	1006.1	62.76	242.5	1022.2
Standard deviation	3.28	13.39	53.63	3.29	12.37	53.77
CV(%)	5.1	5.28	5.33	5.24	5.1	5.26

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