Rat 8-hydroxydeoxyguanosine (8-OHdG) elisa detection kit operation - Database & Sql Blog Articles

Test - Uppercase JPG - Memory 146K

Kaixin Micro Test

Memory Chip eMCP H9TQ64A8GTMCUR-KUM 8+8

STD-MUA-8

Test Probe P100-M3

Rat 8-Hydroxydeoxyguanosine (8-OHdG)

ELISA Test

Kit

Operating Instructions

This kit is for research use only. Not for human or animal diagnostic purposes.

Experimental Principle

The ELISA Kit uses a double-antibody sandwich method to detect the concentration of 8-hydroxydeoxyguanosine (8-OHdG) in rat samples. A microplate is pre-coated with purified rat 8-OHdG antibody, and the sample is added sequentially. The antigen binds to the immobilized antibody, followed by incubation with an HRP-conjugated secondary antibody. After washing, the substrate TMB is added, leading to a color change that is proportional to the amount of 8-OHdG present. The reaction is stopped with an acidic solution, and the absorbance is measured at 450 nm using a microplate reader. The concentration of 8-OHdG in the sample is determined by comparing it to a standard curve.

Kit Composition

1. 130x Washing Solution – 20ml × 1 bottle 2. Stop Solution – 6ml × 1 bottle 3. Enzyme Standard Reagent – 6ml × 1 bottle 4. Standard (200ng/L) – 0.5ml × 1 bottle 5. Enzyme-Labeled Coating Plate – 12 wells × 8 6. Sample Diluent – 6ml × 1 bottle 7. TMB Substrate A – 6ml × 1 bottle 8. TMB Substrate B – 6ml × 1 bottle 9. Standard Dilutions – 1.5ml × 1 bottle 10. Instruction Manual – 1 copy 11. Sealing Film – 2 sheets 12. Sealed Bag – 1

Sample Requirements

1. Samples should be processed as soon as possible after collection. If not tested immediately, store at -20°C. Avoid repeated freeze-thaw cycles. 2. Sodium azide (NaN3) should not be present in the samples, as it may inhibit horseradish peroxidase (HRP) activity.

Testing Procedure

1. Standard Dilution: Prepare a serial dilution of the standard according to the provided table. 2. Loading: Add 50μL of standard or sample diluent to each well, then add 10μL of sample (final dilution 5x). Mix gently. 3. Incubation: Seal the plate and incubate at 37°C for 30 minutes. 4. Washing: Wash 5 times with diluted washing buffer. 5. Enzyme Addition: Add 50μL of enzyme-labeled reagent to each well (except blank). 6. Incubation: Repeat step 3. 7. Washing: Repeat step 4. 8. Color Development: Add 50μL of TMB A and B, incubate at 37°C for 15 minutes. 9. Stop Reaction: Add 50μL of stop solution to each well. 10. Measurement: Read OD450 within 15 minutes using a microplate reader.

Calculation

Plot a standard curve using OD values versus known concentrations. Use linear regression to calculate the unknown sample concentration, and multiply by the dilution factor to obtain the actual value.

Precautions

1. Allow the kit to reach room temperature before use. Store unopened plates in a sealed bag. 2. If washing solution crystallizes, dissolve it in warm water before use. 3. Use a pipette for accuracy; avoid cross-contamination. 4. Always run standards in duplicate. If sample OD exceeds the highest standard, dilute and retest. 5. Use a new sealing film for each test. 6. Keep substrates away from light. 7. Follow all steps strictly. 8. Treat all waste materials as biohazardous. 9. Do not mix reagents from different batches. 10. In case of discrepancy, the English manual takes precedence.

Storage Conditions & Expiration

1. Store at 2–8°C. 2. Shelf life: 6 months from the date of manufacture.