Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit

The ELISA plate follows the standard size of microplate: 127.64 mm x 85.60 mm x 14.22 mm(L x W x H).

Valid for 12 months since the production date. For the shelf life of specific batch number, please check the label printed on the kit. Before delivery, all FineTest® ELISA Kits have been subject to strict quality test.

These information is proprietary. Please contact us to learn more about clonality (polyclonality or monoclonality) and host species.

Not suggested. ELISA reagents are optimized for specific batch.

Yes. The ELISA plate is dismounted. Enough component volumes are offered by 96T ELISA kit, supporting two groups of standard curve.

Used up within 12h.

FineTest® can't support validation of standard concentration outside of standard curve. Ranges of standard curve have been validated among many batches and experimenters, showing stable and accurate performance. The lowest standard concentration is the minimized range for reliable detection results. Adding higher or lower concentration of standard may cause inconsistent signal or false positive.

Matrix components in serum/plasma can affect detection results. Blocking components in sample dilution buffer can decrease or remove the interference. The dilution can reduce the matrix difference between sample and standard to get better accuracy.

FineTest® can't determine the half-life of protein in the sample(e.g. serum, plasma or cell culture supernatant). Usually, it's suggested to detect prepared sample immediately or aliquot sample to refrigerate in a disposable container. Avoid freeze-thaw cycle to prevent protein degradation.

Due to the specificity of each sample, it's hard to forecast and depend on sample preparation as well as analytical characteristics. Please contact us to get detection data for reference.

It depends on whether duplicate assay for your sample is required. Such as, you can detect 80 samples without duplication or 40 samples.

Temperature is the important factor for ELISA binding reaction. The reaction of samples at a consistent temperature requires for equilibrating all reagents to room temperature before the assay, including tested samples. Avoid the inaccurate ELISA assay results caused by temperature differences.

To get more accurate assay results, it's strongly suggested to conduct duplicate assay for the standard and sample. Duplicate assay can calculate average value to ensure more accurate assay results and solve drift caused by misoperation during the assay. Calculation of CV evaluates experimental operation and precision of the ELISA kit.

Follow the suggested method in the manual to store the standard. Before dissolving the standard, transient centrifugation is required to completely collect the powder. Confirm the fully dissolution and mixing of the standard(about 10min). Then, conduct steps of series dilution. Ensure the fully mixing and accurate pipetting in each step. Properly stop the staining. Choose suitable fitting equation to plot the standard curve.

To keep the consistent environmental condition of all plates, stacking is not suggested during incubation.

Some proteins or analytes can bind with glass or polystyrene. However, they are not easy to bind with polypropylene tube.

ELISA assay finally requires for enzyme-catalyzed substrate to complete the staining reaction. Stopping the reaction at the best time is the important factor for successful ELISA assay. When the blue complex appears after adding TMB substrate for 12min, read the O.D. absorbance at 620nm. The staining can be stopped, when the OD value of the darkest color at 620nm is between 0.8 - 0.9. The relevant OD value at 450nm for stopping the staining is between 2.0 - 2.5. If the OD value at 620nm is lower than 0.8, the staining duration can be properly extended.

Dual wavelength for reading in ELISA assay mainly aims to remove non-specific background interference, and improve accuracy and reliability of the detection. FineTest® usually suggests to set the corrected wavelength as 570nm or 630nm in a microplate reader. The OD value may be higher when directly reading at 450nm. However, the accuracy is lower.

FineTest® recommends to use ELISA curve fitting software and validate ELISA kit with 4 - pl curve fitting. 4 - pl curve fitting mainly aims to accurately describe the nonlinear dynamic characteristics of antigen - antibody binding. Thus, the quantitative accuracy of low or high concentration of sample is improved. Experimental errors are decreased. The reliability and reproducibility of assay results are ensured.

Dilute the sample with dilution buffer and conduct the detection again. Ensure the detected value of the sample falls in the range of standard curve.

If you published paper using FineTest® ELISA Kits, you will obtain US$150-US$550 coupon, please contact us for more details.