Description

This kit is able to detect 6 rat antibody heavy chain isotypes and 2 light chain subclasses (IgG1, IgG2a, IgG2b, IgG2c, IgA, IgM, kappa, and lambda) within 5 minutes, fast and convenient; an improvement over the traditional way: ELISA. This kit is shaped like a cassette. After loading 100 µL of solution, a dark line will appear showing the isotype of the antibody. Another line will appear on the control (C), showing a successful test.

Cat#: ISO-R8-20

Tests: 20

Benefit:

The Rapid Rat Immunoglobulin Isotyping Kit is a five-minute lateral flow assay with ELISA sensitivity for monoclonal antibody class and subclass determination. It can determine 6 rat antibody heavy chain isotypes and 2 light chain subclasses (IgG1, IgG2a, IgG2b, IgG2c, IgA, IgM, kappa, and lambda).

Acceptable Sample Types:

Hybridoma culture supernatant and purified monoclonal antibody are suitable samples. But 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.

Sample Requirements and Dilution Conditions:

Cell Culture Supernatant: Recommended dilution is 1:10~1:100 in PBS.

Purified Antibody: Dilute samples to 0.5 – 2 µg/mL in PBS.

Ascites: Ascites fluid is not recommended since it usually contains multiple Igs. However, this kit can still be used to determine the relative distribution of the main isotypes in a sample of ascites fluid. The recommended dilution is 1:10,000 in PBS.

Kit Components:

The isotyping kit has three different sizes with 5, 10 or 20 pouches containing two cassettes per pouch: one cassette is for determining IgG1, IgG2a, IgG2b, IgG2c and the other cassette is for determining Kappa, Lambda, IgA and IgM. Both strips contain a control line.

Protocol:

1) Place the cassettes in horizontal surface.

2) Add 80-100 µL antibody solution directly to the Sample Well. Wait for 5 minutes.

3) A red test line at the location corresponding to antibody-specific isotype and a red control line at “C’ position will appear within 1-5 minutes.

NOTE:

1) Do not read after 20 minutes.

2) If the color of tissue culture medium causes high background, 40-60 µL PBS can be chased to clear the background. 3) Promptly return unused strips in their original bag and keep dry.

Result Interpretation:

1. Invalid result: no red line at “C”.

2. Negative result: only one red line at “C”.

3. Positive results:

a). Single monoclonal antibody: In addition to the positive control “C” line, only one heavy chain and one light chain will appear.

b). Multiple monoclonal antibodies: In addition to the positive control line, more than one heavy chain and/or one light chain will appear. The line intensity can be used to judge the dominant isotypes.

Performance:

Minimum detectable concentration is 100 ng/mL. There is no cross-reactivity among each isotype.

Storage:

Store the strips in their original packaging at room temperature. This product is stable for up to 18 months from the certified date. Freezing the strips is not recommended.