The Five-Minute Rat mAb Isotyping Kit provides a lateral flow assay for monoclonal antibody class and subclass, and light chain type determination. When a sample is loaded onto the cassette, the antibodies will form soluble complexes with pre-embedded gold conjugates. These complexes are resolved and captured by class or isotype specific antibodies impregnated at different areas of the membrane. As a result, a red line will appear at the corresponding region with isotype indicated.
The kit contains 5 (Cat. E001-K003-5) or 20 (Cat. E001-K003-20) pouches with 2 cassettes in each pouch. One cassette is for assessing IgG1, IgG2a, IgG2b, IgG2c and the other one is for Kappa, Lambda, IgA and IgM. Please note that a control ("C") line is included in both cassettes.
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.