Ig subclass/identification/typing kit instruction manual

Mouse monoclonal antibody Ig/subclass identification ELISA kit instruction manual

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I. Principle of reagent: This kit uses the double antibody sandwich method to identify the class and subclass of monoclonal antibody or specific affinity purified monoclonal antibody in mouse lymphocyte hybridoma culture supernatant (can distinguish IgG1\IgG2a\IgG2b \IgG3\IgM\IgA). The secondary antibody coated with the mouse antibody consensus site is coated with a microplate, and combined with the mouse antibody in the culture supernatant added, and then HRP-labeled anti-mouse various and sub-class antibodies are reacted, respectively, and finally used. The TMB substrate system is developed and terminated with dilute sulfuric acid, and the absorbance is measured by a microplate reader to determine the class or subclass of the monoclonal antibody to be tested. With a high resolution, this kit is a reliable tool for identifying mouse monoclonal antibodies and subclasses.

Second, the composition:

Third, the scope of use: for the identification of mouse monoclonal antibody Ig, subclass and related content.

Fourth, the use of:

1, the first kit brought to room temperature (about 30 minutes), and then with pure water cleaning solution formulated working concentrations (a concentrated cleaning solution plus 19 parts of pure water).

2, taken microtiter plates. Each specimen requires 6 wells and the positive control has 6 wells. Excess storage in a ziplock bag, remember to put a desiccant.

3, the cells (or specific affinity purified antibody) ELISA microplate were added 50 l culture supernatant was added to each sample aperture 6, the positive control is added per well of a 6-well 50μl. Then add 50 μl of the sample dilution to each well. The plate was attached and incubated at 37 ° C for 30 minutes.

4 , discard the liquid in the plate, wash the plate 5 times and pat dry or machine wash 5 times on the absorbent material without fibers. Then, 6 μl of each of the enzyme-labeled secondary antibodies were added to each of the 6 wells of each specimen, and the same was used for the 6 wells of the universal positive control. Mark on the sample image or on the plate. The plate was attached and incubated at 37 ° C for 30 minutes.

5 , aspirate the liquid in the plate, wash the plate 5 times and pat dry or machine wash 5 times on the absorbent material without fibers. 50 μl of each of the developer A and the developer B was added to each well, and a new sealing film was applied for 20 minutes in the dark at 37 ° C.

6, this kit specifically good results can be observed visually in general, see the blue color hole corresponding anti-HRP II knows that the specimen Ig class or subclass. The reaction stop solution (50 μl per well) can be terminated, and the wavelength is measured by a microplate reader at 450 nm, and the 630 nm reference wavelength is measured by dual-wavelength. See the high-value well corresponding to the enzyme-labeled secondary antibody to know the Ig class or subclass of the sample. class. The positive control 0D is less than 0.5. The experiment is invalid and needs to be redone.

Five, product features: high sensitivity, high specificity, easy to determine results. Of course, if you feel that a lot of reagents can be prepared to save a fee, you can go to the website and choose the simple antibody identification product C030215 that we have prepared for you. Of course, you can also hand over the specimen to us next time and will not increase the cost! !

Sixth, preservation conditions: 2-8 °C from the light preservation period of one year. Improper storage or too frequent opening may result in a shorter period of validity.

Seven, note:

1, although a long time after the expiration of this kit as well as use value but still you use in the period.

2 , in the detection of ascites monoclonal antibody to be diluted to 1 / 50,000, although it can be distinguished but it is not recommended. The composition of such samples is very complex and has the potential to interfere with the results. In this case, you can choose our C030215 antibody identification reagent. It will bring you satisfactory results.

3 , when you wash the board, you must fill the hole, stay for 10 seconds and then discard it, and finally take a clean shot. In particular, when the plate is washed after the enzyme-labeled secondary antibody is incubated, the enzyme must be sucked out rather than scooped out. This is important for the results when washing the plates by hand.

4 , the board that has not been used up once should be sealed with a desiccant and sealed with the box.

5 , do not violently shake when putting the board, so as to avoid cross-contamination between the holes, the wrong result.

6, abnormal results, please contact us at any time.

Special note: Persons who do not have the basic knowledge and operation of ELISA should not complete this or similar experiments, as this may bring you wrong results and unnecessary losses.