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Mouse Immunoglobulin Isotyping ELISA Kit (MOFI01463)

SKU:
MOFI01463
Product Type:
ELISA Kit
Size:
96 Assays
ELISA Type:
Qualitative ELISA
Synonyms:
monoclonal
Reactivity:
Mouse
€599
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Description

system_update_altDatasheetsystem_update_altMSDS
Product Name: Mouse Immunoglobulin Isotyping ELISA Kit (MOFI01463)
Product Code: MOFI01463
Size: 96 Assays
Alias: Mouse Immunoglobulin Isotyping ELISA
Detection Method: Qualitative ELISA
Application: 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.

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