Human Caspase-1 p20(Caspase-1 p20) ELISA Kit

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 Caspase-1 p20 antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with Caspase-1 p20 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 Caspase-1 p20 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Product Name

Human Caspase-1 p20(Caspase-1 p20) ELISA Kit

Alias

Caspase-1 p20 ELISA Kit, CASP-1 ELISA Kit, Caspase-1 ELISA Kit

Catalogue No.

EH5100

Size

48T/96T

Species

Human

UniProt ID

P29466

Sample Type

Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples

Detection Method

Sandwich ELISA, Double Antibody

Detection Wavelength

OD450

Reaction Duration

4 hours

Range

31.25-2000pg/ml

Sensitivity

18.75pg/ml

Storage

2-8°C(Sealed), Don't cryopreserve.

Specificity

Specifically binds with Caspase-1 p20 , 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
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

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: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.

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	0.08	0.082	0.081
0.488	0.173	0.179	0.176
1.953	0.246	0.254	0.25
7.813	0.434	0.446	0.44
31.25	0.79	0.812	0.801
125	1.043	1.073	1.058
500	1.643	1.691	1.667
2000	2.244	2.31	2.277

Recovery

Add a certain amount of Caspase-1 p20 into the sample. Calculate the recovery by comparing the measured value with the expected amount of Caspase-1 p20 in the sample.

Sample Type	Recovery Range(%)	Average(%)
serum(n=10)	85-104	92
EDTA plasma(n=10)	85-100	91
Heparin plasma(n=10)	85-105	100

Linearity

Dilute the sample with a certain amount of Caspase-1 p20 at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type	1:2	1:4	1:8
serum(n=10)	88-101%	85-96%	91-96%
EDTA plasma(n=10)	88-105%	85-92%	86-98%
Heparin plasma(n=10)	86-105%	86-99%	90-98%

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)	58.45	251.4	976.1	59.4	242.9	986.3
Standard deviation	3.07	12.22	47.24	3.04	12.36	52.67
CV(%)	5.26	4.86	4.84	5.12	5.09	5.34

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