The Five-Minute Hamster mAb Isotyping Kit is a five-minute lateral flow assay with ELISA sensitivity for monoclonal antibody isotype determination. Acceptable sample types include hybridoma culture supernatant and purified monoclonal antibody. Ascites fluid is not recommended since it usually contains multiple immunoglobulins. However, this kit can still be used to determine the relative distribution of the main isotypes in a sample of ascites fluid.
The kit contains 5 (Cat. E001-K006-5) or 20 (Cat. E001-K006-20) pouches with 2 same cassettes in each pouch. The cassette is for assessing IgG1, IgG2, IgG3, and Kappa. Please note that a control (“C”) line is included in both cassette.
The minimum detectable concentration is 100 ng/mL. Below is our recommendation on sample preparation.
|Cell Culture Supernatant||Purified Antibody||Ascites|
|1:100 dilution in PBS (optional)||0.5-2 µg/mL in PBS||1:10,000 in PBS *|
* Normally, ascites fluid contains multiple Igs. The kit can only be used to determine the relative distribution of the main isotypes.
Please keep the strips in their original packaging at room temperature. This product is stable for up to 18 months from the certified date. Please do NOT freeze the strips.
1.Place the cassettes on a flat surface.
2.Add 100 µL sample directly to the well and wait for 5 minutes.
3.A red line should appear on the membrane with specific isotypes indicated at the same position. A red line should also appear at the “C” position.
NOTE1:Prepare samples before removing the strips from the sealed pouch.
NOTE2:Do not read after 20 minutes.
NOTE3:The color of tissue culture medium may cause high background. To address this issue, additional 40-60 µL PBS can be added into the well to clear the background.
NOTE4:The reactivity for Igs from the two major hamster species is shown in the following table.