ultrasensitive ELISA and ImmunoPCR

Immuno-PCR (iPCR) is a powerful method for detecting ultra-low quantities of antigens. It combines the advantages of both enzyme-linked immunosorbent assay (ELISA) and PCR in specificity, sensitivity and easy to adapt. Despite its potential, iPCR is an underutilized method as evidenced by the low number of publications on its routine application and unavailability of validated, ready-to-use commercial kits. To make it possible for the researchers to detect various low-abundant analytes, Novatein Biosciences is introducing validated and ready-to-use iPCR conversion kit(cat# NB-iPCR42)and iPCR kits.

Fig. 1. Schematic of iPCR technique

Novatein Bioscience's iPCR conversion kit (cat# NB-iPCR42) makes it possible to convert moderately sensitive conventional ELISA to an ultra-sensitive iPCR. The kit has all the reagents except antibodies and standard. One of the main advantages of using Novatein Bioscience's iPCR conversion kit is its scalability to high-throughput systems. In a typical work day, ~500 samples can be quantified in triplicate.

Other advantages of iPCR over conventional ELISA include:

• 1.10-100 fold improvement in the assay range making it possible to quantify low-abundant targets like HMGB1, HCP etc
• 2.Low amounts of sample is needed, which means saving of precious samples and no matrix effect.

In addition, Novatein Biosciences is offering the following validated iPCR kits:

• Human cytokine iPCR kit
• Mouse cytokine iPCR kit
• Other species cytokine iPCR kit
• Host Cell Protein (HCP) iPCR Kit

Customized antibody-DNA  conjugate preparation service

(please contact info @ novateinbio.com for detail)

1. Plate Coating: Coat strips with Capture Antibody overnight at 2 -8 0C. 3x washes.
2. Blocking: Wash strips, block with Buffer B overnight at 2 -8 0C. 3x washes. Air dry and strips can be stored at 2 -8 0C for weeks to months desiccated for future use.

1. Antigen/protein incubation: Wash strips, make  standard dilution in Buffer B, do cell lysate dilution from >= 5-fold in Buffer B, incubate overnight at 2-8°C, or 2-hr incubation at room temperature. 6 x washes.
2. Detection AB-DNA complex incubation: dilute the Antibody-DNA complex at 1:100 to 1:200 dilution, add to designated wells and incubate for 45 – 60 min, 10x washes.
3. qPCR:  Mix PCR master mix with Primer-probe mix in nuclease-free water, dispense into strip wells, stir for 1 min at 600 rpm at RT. Transfer to PCR machine, seal the wells and start the 40-cycle PCR. After1.5 hours, analyze results.