F-TESTO(Free Testosterone) ELISA Kit (E-EL-0165)

Q1:I want to measure the content of testosterone in the supernatant of cell culture. Does the culture medium containing phenol red affect the detection of testosterone?

The medium containing phenol red may have an effect on the detection of testosterone. Phenol red is used in the medium as an indicator of PH: red when neutral, yellow when acidic, and purple when alkaline. Studies have shown that phenol red can mimic the effects of sterol hormones (especially estrogen). To avoid sterol reactions, cultured cells, especially mammalian cells, are cultured without phenol red. Introduce a tip: when the medium is packaged, leave a tube with phenol red and put it in the incubator to balance (you can see the change in PH value, and you can see whether the medium is polluted). Put in the incubator before culture for balanced use (can avoid the use of phenol red during culture).

Q2:Why is it necessary to add a protease inhibitor in tissue sample preparation during an Elisa experiment? Will it affect the detection significantly if there is no protease inhibitor?

Tissue samples may contain endogenous or exogenous proteases during processing, leading to degradation of extracted proteins. Therefore, it's necessary to add protease inhibitors during processing to ensure the integrity of target proteins. If customers can keep samples cold and handle them quickly during processing, omitting the protease inhibitor may not have a significant effect. After preparation, samples should be tested promptly or immediately aliquoted and frozen at -20°C or -80°C.

Q3:Which variant of the spike protein does this kit detect? Do you have a kit specific to detect the spike protein for the omicron variant?

This kit is designed for the original strain of the new crown virus, and the omicron variant has not been verified. However, we have verified 26 recombinant variants of the SARS-CoV-2 spike protein through the kit. For more information, customers can refer to the kit instructions (https://file.elabscience.com/Manual/covid_19/E-EL-E605 .pdf).

Q4:What is the range of enzyme activity of your IL-2 freeze-dried powder

Currently our freeze-dried powder is a concentration unit with no information on the activity unit for the time being.

Q5:What is the principle of adding stop solution to stop color reaction in ELISA experiment?

On the one hand, the activity of HRP enzyme is destroyed and the catalytic function of HRP enzyme is lost. On the other hand, the final color changes due to a change in pH.

Q6:The absorbance of the cell supernatant is less than that of the sample diluent alone, so how to concentrate the sample?

The amount of medium can be reduced for subsequent drug administration and modeling.

Q7:My sample volume is small. Can I reduce some reagents proportionally?

No, the detection system of our kits requires strict adherence to the specified sample volume to ensure accurate detection. If the sample volume is insufficient, consider diluting appropriately, but first conduct a pre-experiment to confirm the suitable dilution factor.

Q8:May I ask which type of plate to choose when ELISA testing?

According to the different bottom, it is divided into flat bottom, U-shaped bottom, V-shaped bottom and so on. The index of refraction of the flat bottom is low, which is suitable for detection in the enzyme reader. According to the color can be divided into transparent, black, white. Transparent is commonly used for the most general enzyme-linked immunoassay.

Q9:Is the TGF-β1 ELISA Kit detecting the active form of TGF-β1 or the precursor?

The active TGF-β dimer was detected

Q10:I need to measure corticosterone and testosterone in hair samples. Are there any suggested sample extraction methods?

Hair sample preparation method: Wash hair samples with methanol by adding 5 mL of HPLC-grade methanol to each sample, rotating for 3 minutes, then decanting excess methanol and rinsing hair twice. After washing, place the hair samples on aluminum foil, dry for 3 days in a protective cap. Weigh the dried hair samples and transfer them to 2ml polypropylene tubes containing stainless steel grinding beads. Place the tubes containing hair and beads in a bead beater, grind each sample for 2 minutes to produce powder. After grinding, add 1.5 mL of methanol to the tubes containing hair powder. Rotate samples slowly for 24 hours to extract steroids. Centrifuge at 10000 g for 4 minutes, transfer 0.6 mL of methanol supernatant containing steroids to new 1.5 mL microcentrifuge tubes. Dry the samples in a protective hood for 2-3 days to evaporate the methanol. Dilute the dried extract with 0.4 mL dilution buffer from the kit for detection.