Products
Rat Eln (Elastin) QuickTest ELISA Kit
This kit was based on sandwich ELISA method. The experiment lasted 120 minutes. Capture antibody was conjugated to an affinity tag that was recognized by a specific antibody coated on the QuickTest plate. Add the Cap/Det Ab working solution into each well, then add the standards and pilot samples into individual wells. If the sample contains ELN, a capture antibody-ELN-biotin-detection antibody complex was formed. After incubation, unbound conjugates were removed by wash buffer. HRP-Streptavidin was added. After washing, TMB substrates were added to visualize HRP enzymatic reaction. 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. The concentration of ELN in the sample was calculated by drawing a standard curve. The concentration of the target substance is proportional to the OD450 value.
- Product Name
- Rat Eln (Elastin) QuickTest ELISA Kit
- Alias
- ELN ELISA Kit, Elastin ELISA Kit, Tropoelastin ELISA Kit
- Catalogue No.
- QT-ER0508
- Size
- 48T/96T
- Species
- Rat
- UniProt No.
- Q99372
- Sample Type
- Serum, plasma, cell culture supernatant, cell lysate or tissue lysate, other biological fluid samples
- Detection Method
- Sandwich ELISA, Double Antibody
- Detection Wavelength
- OD450
- Reaction Duration
- 120 minutes
- Range
- 0.625-40ng/ml
- Sensitivity
- 0.375ng/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with ELN , 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
-
- 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: 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.
STD.(ng/ml) OD-1 OD-2 Average 0 0.075 0.077 0.076 0.625 0.173 0.171 0.173 1.25 0.241 0.235 0.238 2.5 0.436 0.433 0.418 5 0.711 0.705 0.674 10 1.005 1.012 1.034 20 1.659 1.675 1.631 40 2.112 2.061 2.076 
- Recovery
-
Add a certain amount of ELN into the sample. Calculate the recovery by comparing the measured value with the expected amount of ELN in the sample.
Sample Type Recovery Range(%) Average(%) serum(n=10) 88-104 98 EDTA plasma(n=10) 86-98 91 Heparin plasma(n=10) 87-105 96 - Linearity
-
Dilute the sample with a certain amount of ELN at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type 1:2 1:4 1:8 serum(n=10) 88-104% 85-101% 87-104% EDTA plasma(n=10) 82-89% 84-99% 82-98% Heparin plasma(n=10) 80-96% 85-100% 80-99% - Precision(%)
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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) 1.23 5.26 20.89 1.26 5.04 18.79 Standard deviation 0.05 0.17 0.75 0.05 0.19 0.73 CV(%) 4.22 3.21 3.61 3.84 3.82 3.89 - 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