This Human DEFB4 ELISA Kit was designed for the quantitative measurement of Human DEFB4 protein in serum, urine, cell culture supernates, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 62.5 pg/mL-4000 pg/mL and the sensitivity is 15.6 pg/mL.
||defensin, beta 4
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||Homo sapiens (Human)
||serum, urine, cell culture supernates, tissue homogenates
||62.5 pg/mL-4000 pg/mL
|Intra-assay Precision (Precision within an assay): CV%<8%|| || || |
|Three samples of known concentration were tested twenty times on one plate to assess. || |
|Inter-assay Precision (Precision between assays): CV%<10%|| || || |
|Three samples of known concentration were tested in twenty assays to assess. || || |
| || || || || || || |
|To assess the linearity of the assay, samples were spiked with high concentrations of human β-defensins 2 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.|
| ||Sample||Serum(n=4)|| |
|1:1||Average %||87|| |
|Range %||81-92|| |
|1:2||Average %||94|| |
|Range %||90-100|| |
|1:4||Average %||98|| |
|Range %||91-110|| |
|1:8||Average %||91|| |
|Range %||85-98|| |
|The recovery of human β-defensins 2 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.|
|Sample Type||Average % Recovery||Range|| |
|Serum (n=5) ||93||89-98|| |
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| || || || || || || |
|These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
|4000||2.821 ||2.752 ||2.787 ||2.689 || |
|2000||2.182 ||2.076 ||2.129 ||2.031 || |
|1000||1.438 ||1.364 ||1.401 ||1.303 || |
|500||0.802 ||0.793 ||0.798 ||0.700 || |
|250||0.471 ||0.437 ||0.454 ||0.356 || |
|125||0.302 ||0.290 ||0.296 ||0.198 || |
|62.5||0.243 ||0.229 ||0.236 ||0.138 || |
|0||0.099 ||0.097 ||0.098 || || |
- A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human DEFB4 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled DEFB4 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
- One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- An instruction manual
|Materials not provided
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Test tubes for dilution
|ELISA Data Analysis
||Watch ELISA data processing video & download Curve Expert if needed
|ELISA kit FAQs
||Store at 2-8°C. Please refer to protocol.
||3-5 working days