The Mouse Monoclonal Antibody Isotype ELISA Kit (MOFI01463) is a highly sensitive and specific kit designed for the accurate detection of mouse monoclonal antibody isotypes in various biological samples. This kit is ideal for researchers studying immune responses and antibody production in mice, as it provides reliable and reproducible results.Mouse monoclonal antibody isotypes play a crucial role in immune responses and can provide valuable information about the immune system's reaction to pathogens and diseases.
By accurately measuring these isotypes, researchers can gain insight into the effectiveness of experimental treatments and vaccines, as well as monitor the progression of immune responses in mouse models.With its high sensitivity and specificity, the Mouse Monoclonal Antibody Isotype ELISA Kit (MOFI01463) is a valuable tool for a wide range of research applications in immunology, infectious diseases, and vaccine development. Trust AssayGenie for accurate and reliable results in your antibody research.
For the qualitative isotype determination of mouse immunoglobulins (IgG1, IgG2a, IgG2b, IgG3, and IgM) from hybridoma cell culture supernatant or purified antibodies.
Storage:
2-8°C for 6 months
Note:
For Research Use Only
Intra Assay:
CV <8%
Inter Assay:
CV <10%
Component
Quantity
Storage
ELISA Microplate(Dismountable)
8-12 strips
-20°C
Negative Control
1 mL
2-8°C
Positive Control
1 mL
2-8°C
Sample Dilution Buffer
20 mL
2-8°C
HRP-Streptavidin Conjugate(SABC)
60ul/120ul
2-8°C (Avoid Direct Light)
Wash Buffer (25X)
30 mL
2-8°C
Antibody dilution buffer
5 mL/10 mL
2-8°C
TMB substrate
5ml/10ml
2-8°C (Avoid Direct Light)
Plate Sealer
5
-
Other materials and equipment required:
Microplate reader with 450 nm wavelength filter
37°C incubator
Automated plate washer
Precision single and multi-channel pipette and disposable tips
Clean tubes and Eppendorf tubes
Deionized or distilled water
*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Step
Procedure
1.
Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly.
2.
Set Positive, negative control and test samples wells on the pre-coated plate respectively. One test sample need 5 wells, 5 positive and 5 negative control wells. If need to detect one sample A, you need 1×5 (remark number as A1,A2,A3,A4,A5)+10=15wells. If need to detect two samples, you need 2×5+10=20 wells, and so on. And then, record their positions.
3.
Add 100μl of positive and negative control working solution into the control wells.
4.
Seal the plate with a cover and incubate at 37°C for 90 min.
5.
Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Wash plate 2 times. Do NOT let the wells completely dry at any time.
6.
Add 100μl of biotin labeled antibody working solution to the corresponding wells (e.g., add Biotin-anti-mouse IgG1 antibody to A1,B1,C1 and the corresponding negative and positive control Wells; Add biotin-anti-mouse IgG2A to A2,B2,C2, and corresponding negative and positive control wells, and so on).
7.
Seal the plate with a cover and incubate at 37°C for 30 mins.
8.
Remove the cover, and wash plate 3 times with Wash buffer. Add 100μl of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
9.
Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
10.
Add 90μl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-15 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.)
11.
Add 50μl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. Read the OD. Absorbance at 450 nm in a microplate reader immediately after adding the stop solution.
