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**Human Mann's Split-Head Antibody (SM Ab) Enzyme-Linked Immunoassay**
**Purpose:**
This ELISA kit is designed to detect the presence of Human Mann's Split-Head Antibody (SM Ab) in human serum, plasma, and other related biological samples. It provides a reliable method for screening and quantifying SM Ab levels, which may be useful in clinical diagnostics or research settings.
**Experimental Principle:**
The assay is based on the double-antigen sandwich immunoassay technique. A solid-phase antigen is pre-coated onto microtiter plate wells. The sample is added, allowing SM Ab present in the specimen to bind to the immobilized antigen. After washing to remove unbound components, an HRP-conjugated secondary antigen is introduced, forming a complex of antigen-antibody-enzyme-labeled antigen. Following another wash, TMB substrate is added, and the reaction is stopped with a stop solution. The color change, from blue to yellow, is measured at 450 nm using a microplate reader. The OD value is compared to a cutoff value to determine the presence or absence of SM Ab in the sample.
**Sample Preparation:**
It is recommended to process the samples as soon as possible after collection. If immediate testing is not feasible, store the samples at -20°C. Avoid repeated freeze-thaw cycles. Note that samples containing sodium azide (NaN3) should not be used, as it may inhibit horseradish peroxidase (HRP) activity, leading to inaccurate results.
**Procedure Overview:**
1. **Labeling:** Assign unique numbers to each well on the microplate. Include two negative control wells, two positive control wells, and one blank control well.
2. **Loading:** Add 50 µL of negative and positive controls to their respective wells. For the test samples, add 40 µL of diluent followed by 10 µL of the sample into each sample well. Ensure the sample is added to the bottom of the well without touching the sides.
3. **Incubation:** Cover the plate and incubate at 37°C for 30 minutes to allow antibody-antigen binding.
4. **Washing:** Prepare a 30x diluted washing solution with distilled water. Wash the plate five times, ensuring all unbound material is removed.
5. **Enzyme Addition:** Add 50 µL of HRP-conjugated reagent to each well except the blank control.
6. **Second Incubation:** Repeat the incubation step at 37°C for 30 minutes.
7. **Final Washing:** Perform the same washing procedure as before.
8. **Color Development:** Add 50 µL of TMB A and 50 µL of TMB B to each well. Mix gently and incubate at 37°C for 15 minutes in the dark.
9. **Stop Reaction:** Add 50 µL of stop solution to each well to terminate the reaction. The color will turn from blue to yellow.
10. **Reading:** Measure the absorbance at 450 nm within 15 minutes of adding the stop solution.
**Data Interpretation:**
- **Positive Control:** Average OD ≥ 1.00
- **Negative Control:** Average OD ≤ 0.10
- **Cutoff Value:** Cutoff = Negative Control Average + 0.15
If the sample OD value is above the cutoff, it is considered positive for SM Ab; otherwise, it is negative.
**Note:** This kit is intended for research use only. Proper handling and interpretation are essential for accurate results. Always refer to the manufacturer’s instructions for detailed procedures and safety information.
**Download:**
[Human Mann's Schizophrenia Antibody (SM Ab) Kit Instructions](#)
*Click to download the complete instruction manual.*
**Company Information:**
For more details, visit the official booth of **Shanghai Hengyuan Biotechnology Co., Ltd.**
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