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 IL-6 antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with IL-6 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 IL-6 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Manual- Product Name
- Goat IL-6(Interleukin 6) ELISA Kit
- Alias
- Interleukin-6 ELISA Kit, IL-6 ELISA Kit, B-cell stimulatory factor 2 ELISA Kit, BSF-2 ELISA Kit, CTL differentiation factor ELISA Kit, CDF ELISA Kit, Hybridoma growth factor ELISA Kit, Interferon beta-2 ELISA Kit, IFN-beta-2 ELISA Kit, IL6 ELISA Kit, IFNB2 ELISA Kit
- Catalogue No.
- EG0029
- Size
- 48T/96T
- Species
- Goat
- 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
- 15.625-1000pg/ml
- Sensitivity
- 9.375pg/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with IL-6 , 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 0 0.11 0.114 0.112 15.625 0.15 0.154 0.152 31.25 0.185 0.191 0.188 62.5 0.294 0.302 0.298 125 0.489 0.503 0.496 250 0.736 0.758 0.747 500 1.441 1.483 1.462 1000 2.264 2.33 2.297 - Recovery
-
Add a certain amount of IL-6 into the sample. Calculate the recovery by comparing the measured value with the expected amount of IL-6 in the sample.
Sample Type Recovery Range(%) Average(%) serum(n=10) 91-105 99 EDTA plasma(n=10) 88-102 95 Heparin plasma(n=10) 85-104 96 - Linearity
-
Dilute the sample with a certain amount of IL-6 at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type 1:2 1:4 1:8 serum(n=10) 87-104% 85-98% 85-97% EDTA plasma(n=10) 91-105% 83-101% 80-92% Heparin plasma(n=10) 93-103% 83-96% 80-97% - 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) 31.11 126.7 520 31.64 130.1 506 Standard deviation 1.55 6.87 27.82 1.57 6.87 24.9 CV(%) 4.99 5.42 5.35 4.96 5.28 4.92 - 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