Human TIP49A ELISA Kit

From Assay Genie, this Sandwich ELISA kit is designed for the quantitative measurement of TIP49A in human samples. The assay uses a double-antibody sandwich format with colorimetric detection, delivering a sensitivity of 0.188 ng/mL across a working range of 0.313–20 ng/mL. It is intended for use with serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples, and the kit components are stored at 2-8°C for up to 12 months. TIP49A (UniProt Q9Y265, also known as RUVBL1, Pontin 52 and related synonyms) is a protein involved in DNA repair, gene regulation and cell growth, with research applications in the cell cycle area and as a biomarker in conditions such as cancer and neurodegenerative disorders.

The assay procedure follows a standard biotin–streptavidin–HRP workflow. After reagent equilibration and plate setup, standards, samples and blanks are incubated at 37°C for 90 minutes for primary antigen binding, followed by a biotin-labelled detection antibody incubation at 37°C for 60 minutes and an HRP-Streptavidin (SABC) incubation at 37°C for 30 minutes. TMB substrate is then added and incubated in the dark for 10–20 minutes before the reaction is stopped and absorbance is read at 450 nm. Sample preparation guidance includes clotting and centrifugation of serum at 1000 × g for 20 minutes, collection of plasma in anticoagulant tubes with centrifugation at 1000 × g for 15 minutes at 2–8°C, homogenisation of tissue in PBS with protease inhibitors, and clarification of cell culture supernatant at 2500 rpm for 5 minutes.

Reported linearity at 1:2, 1:4 and 1:8 dilutions falls within 88–104%, 84–99% and 89–99% for serum, 87–98%, 82–92% and 82–93% for EDTA plasma, and 88–103%, 83–98% and 81–93% for heparin plasma (n = 5). Spike-and-recovery performance is reported as 85–103% with an average of 96% for serum, 92–105% with an average of 99% for EDTA plasma, and 94–102% with an average of 97% for heparin plasma (n = 5). The pre-coated ELISA microplate, lyophilised standard, biotin-labelled detection antibody, HRP-Streptavidin conjugate, TMB substrate, sample, antibody and SABC dilution buffers, stop solution, wash buffer, plate sealer and technical manual are all included to support a complete run.