- Product Name
- Human CAF(c-terminal fragemnte of agrin) ELISA Kit
- Alias
- CAF ELISA Kit, c-terminal fragment of agrin ELISA Kit
- Catalogue No.
- EH4820
- Size
- 48T/96T
- Species
- Human
- UniProt No.
- O00468
- 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
- 46.875-3000pg/ml
- Sensitivity
- 28.125pg/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with CAF , 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
-
- 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 (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) 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
- 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 | Corrected |
0 | 0.087 | 0.092 | 0.089 | 0.003 |
46.875 | 0.339 | 0.353 | 0.346 | 0.257 |
93.75 | 0.486 | 0.506 | 0.496 | 0.407 |
187.5 | 0.767 | 0.799 | 0.783 | 0.694 |
375 | 1.047 | 1.089 | 1.068 | 0.979 |
750 | 1.475 | 1.535 | 1.505 | 1.416 |
1500 | 1.751 | 1.823 | 1.787 | 1.698 |
3000 | 2.18 | 2.268 | 2.224 | 2.135 |
- Recovery
-
Add a certain amount of CAF into the sample. Calculate the recovery by comparing the measured value with the expected amount of CAF in the sample.
Sample Type | Recovery Range(%) | Average(%) |
serum(n=10) | 92-104 | 92 |
EDTA plasma(n=10) | 92-100 | 93 |
Heparin plasma(n=10) | 90-105 | 97 |
- Linearity
-
Dilute the sample with a certain amount of CAF at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type | 1:2 | 1:4 | 1:8 |
serum(n=10) | 88-102% | 85-104% | 86-102% |
EDTA plasma(n=10) | 86-98% | 85-101% | 82-96% |
Heparin plasma(n=10) | 83-97% | 85-100% | 80-95% |
- 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) | 92.92 | 379.9 | 1468 | 94.6 | 377.8 | 1497 |
Standard deviation | 4.45 | 19.15 | 74.72 | 4.73 | 18.97 | 75.15 |
CV(%) | 4.79 | 5.04 | 5.09 | 5 | 5.02 | 5.02 |
- 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 |