Rapid Mouse Antibody Isotyping kit (Cat# ISO-M6)

Description

Benefit:

The Rapid Mouse Antibody Isotyping Kit (ISO-M6) is a five-minute lateral flow assay with ELISA sensitivity for monoclonal antibody class and subclass determination. It can determine IgG1, IgG2a, IgG2b, IgM, κ, and λ.

Sample Requirements and Dilution Conditions:

Acceptable sample types include hybridoma culture supernatant and purified monoclonal antibody. This kit can also be used to determine the relative distribution and changes of the isotypes in serum under health and disease status.

Cell Culture Supernatant: Depending on the actual titre of the Ab-producing cells or hybridomas, recommended dilution is 1:2~1:50 in PBS.

Purified Antibody: Dilute samples to 0.5 – 2.0 μg/ml in PBS.

Serum: Serum can be diluted 100-500 fold 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, and IgG2b, and the other cassette is for determining Kappa, Lambda and IgM. Both strips contain a control line.

Protocol:

1) Place the cassettes in horizontal plane.
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: Do not read after 20 minutes. If the color of tissue culture medium causes high background, 40-60 μl PBS can be chased to clear the background. Promptly return unused strips in sealed 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: Besides the positive control line, more than one heavy chain and/or light chain will appear. The line intensity can be used to judge the dominant isotypes.

Performance:

Minimal detection limit is 100 ng/ml. There is no cross-reactivity among each isotype.

Storage:

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

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