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 FN1 antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with FN1 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 FN1 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
| Size | |
|---|---|
| 96T | Inquiry |
| 48T | Inquiry |
- Product Name
- Human FN1(Fibronectin) ELISA Kit
- Alias
- FN ELISA Kit, Fibronectin ELISA Kit, FN1 ELISA Kit, CIG ELISA Kit, ED-B ELISA Kit, FINC ELISA Kit, FNZ ELISA Kit, GFND ELISA Kit, GFND2 ELISA Kit, LETS ELISA Kit, MSF ELISA Kit, Cold-insoluble globulin ELISA Kit, CIG) ELISA Kit
- Catalogue No.
- EH0134
- Size
- 48T/96T
- Species
- Human
- UniProt ID
- P02751
- 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
- 1.563-100ng/ml
- Sensitivity
- 0.938ng/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with FN1 , no obvious cross reaction with other analogues.
- Recommended Sample Dilution Ratio
-
The following table shows the recommended dilution ratios for this kit for a limited number of samples for your reference only. (When the sample content is very low, the original solution can be added without dilution, but it is necessary to ensure that the pH is between 6.8 and 8.0, and it does not contain more than 10% organic solvents or high-concentration protein denaturants.)
1. Common sample validation:
Sample Type Recommended Dilution Ratio Content (Please refer to shipped instructions or contact us) (Please refer to shipped instructions or contact us) (Please refer to shipped instructions or contact us) (Please refer to shipped instructions or contact us) (Please refer to shipped instructions or contact us) (Please refer to shipped instructions or contact us) 2. KO sample validation (Detect FN1 KO cells):
Sample Type Dilution Ratio Content Note：ND is lower than the sensitivity of the kit and was not detected
3. Capture antibody and detection antibody by WB KO validation (Detect FN1 KO cells):
- 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.(ng/ml) OD-1 OD-2 Average 0 0.134 0.138 0.136 1.562 0.189 0.195 0.192 3.125 0.24 0.246 0.243 6.25 0.336 0.346 0.341 12.5 0.512 0.526 0.519 25 0.838 0.862 0.85 50 1.328 1.366 1.347 100 2.136 2.198 2.167 
- Recovery
Add a certain amount of FN1 into the sample. Calculate the recovery by comparing the measured value with the expected amount of FN1 in the sample.
Sample Type Recovery Range(%) Average(%) serum(n=10) 85-104 96 EDTA plasma(n=10) 85-98 91 Heparin plasma(n=10) 85-104 98 - Linearity
Dilute the sample with a certain amount of FN1 at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type 1:2 1:4 1:8 serum(n=10) 91-104% 87-103% 87-98% EDTA plasma(n=10) 85-97% 82-101% 82-98% Heparin plasma(n=10) 81-99% 85-98% 85-95% - 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 (ng/ml) 3.03 11.91 47.47 3.08 11.49 48.78 Standard deviation 0.16 0.59 2.43 0.14 0.62 2.38 CV(%) 5.12 4.98 5.12 4.69 5.43 4.87 - 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