Monkey TNF-α culture media OneStep ELISA Kit

Catalog Number: S0C3251 Reactivity: Mk Conjugation: Brand: Starter

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$670 USD

Size: 1x96T

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Product Details

Product Specification

Antigen	TNF-α
Immunogen	Recombinant Protein
Antibody Type	Recombinant mAb
Reactivity	Mk
Purification	Protein A
Stability & Storage	12 months from date of receipt / reconstitution, 2 to 8°C as supplied.

Kit

Precision	Intra-assay: 8.7% Inter-assay: 8.9%
Sample type	Cell culture supernatant
Assay type	Sandwich (quantitative)
Sensitivity	56.24 pg/mL
Range	51.6 pg/mL – 3300 pg/mL
Recovery	Cell culture supernatant: 89%
Assay time	60 minutes

Background

Monkey tumor necrosis factor-alpha (TNF-α), also known as cachectin, is a pleiotropic pro-inflammatory cytokine that plays a central role in host defense and immune regulation. The full-length cynomolgus monkey (Macaca fascicularis) TNF-α precursor consists of 233 amino acids, with a mature secreted form spanning approximately 157 residues. Recombinant monkey TNF-α is typically produced in E. coli as a non-glycosylated polypeptide chain with a molecular weight of approximately 16.7–17.3 kDa. The protein shares high sequence homology with human TNF-α—approximately 98% at the amino acid level for rhesus macaque—which makes monkey models highly relevant for translational research. Biologically active TNF-α exists as a homotrimer and signals through two distinct receptors, TNFR1 and TNFR2, to mediate diverse cellular responses including cytotoxicity, inflammation, and apoptosis. In healthy cynomolgus and rhesus macaques, serum TNF-α levels are generally low, with a reported normal range of approximately 1–10 pg/mL. However, detection of TNF-α in healthy non-human primate serum using multiplex assays has been noted to face challenges regarding recovery and linearity, suggesting that results should be interpreted with caution.

Picture

ELISA

Standard curve
Example of Monkey TNF-α standard curve in Assay Diluent J1.

Spike Recovery
The recovery of Monkey TNF-α was evaluated in activated samples spiked with concentrations spanning the entire assay range.

Spike-and-dilution Linearity
To evaluate assay linearity, culture media sample was spiked with high levels of Monkey TNF-α in different matrices and serially diluted with the corresponding Calibrator Diluent to fall within the assay's dynamic range.

Determination of Minimum Detectable Dose (MDD)
The MDD was determined using three independent lots of assay components. For each lot, 19 replicate measurements of the diluent (zero calibrator) were performed. The mean (AVERAGE) and standard deviation (STDEV) of the 19 replicates were calculated. The MDD for each lot was then calculated according to the following formula:
MDD = 2 × STDEV + AVERAGE

Dilution linearity
The concentrations of Monkey TNF-α were measured and interpolated from the target standard curves and corrected for sample dilution.
The leftmost bar in the graph represents neat biological samples. "Undiluted" was defined as the neat biological sample (100%). The subsequent bars represent two-fold serial dilutions of this reference. The mean target concentration was determined to be 1032 pg/mL in Monkey PBMC.

HOOK Effect Threshold
The upper limit of the HOOK effect was established at 100× the highest calibrator concentration (equivalent to a 2-log10 increase). No HOOK effect was observed below this threshold, confirming that samples within this concentration range yield reliable quantitative results without signal depression.

Intra-Assay Precision (Precision within an assay)
Three samples of known concentration were tested fifteen times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays)
Three samples of known concentration were tested in separate assays to assess inter-assay precision. Assays were performed with at least three lots of components.