Human CD117 molecule (CD117) ELISA kit instruction manual - Database & Sql Blog Articles

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Kit

Can only be used for

scientific

research

, not for medical diagnosis.

Human CD117 molecule (CD117) ELISA kit

User's Guide

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Human CD117 molecule (CD117) ELISA kit

Kit name

Human CD117 molecule (CD117) ELISA kit

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Human CD117 molecule (CD117) ELISA kit

Kit use]

Quantitative detection of CD117 molecule (CD117) in human serum, plasma, and related fluid samples.

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Human CD117 molecule (CD117) ELISA kit

Detection principle

The kit uses a two-antibody sandwich ELISA method. The standard and sample are added to the pre-coated enzyme-labeled plate, incubated for a sufficient time, then washed to remove unbound components. Substrates A and B are added sequentially, and the TMB substrate is converted to a blue product under horseradish peroxidase (HRP) catalysis, turning yellow in acidic conditions. The color intensity is directly proportional to the concentration of CD117 in the sample. OD values are measured at 450 nm, and the sample concentration is calculated based on the standard curve.

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Human CD117 molecule (CD117) ELISA kit

Kit composition

1	Enzyme-labeled coated plate	12 wells × 8 pieces
2	Standard: 200ng/ml	0.6mL
3	Substrate solution A	6mL
4	20× concentrated washing solution	20mL
5	Sample diluent	6mL
6	Enzyme standard reagent	6mL
7	Substrate solution B	6mL
8	Stop solution	6mL
9	Standard dilution	6mL
10	Instruction manual	1 copy
11	Sealing film	1 piece
12	Sealed bag	1 piece

Notes:

Standard should be diluted with standard diluent to concentrations of: 200, 100, 50, 25, 12.5, 6.25 ng/ml.

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Human CD117 molecule (CD117) ELISA kit

Reagents and equipment needed but not provided

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1. 37°C incubator

2. Microplate reader

3. Precision pipettes and disposable tips

4. Distilled water

5. Disposable test tubes

6. Absorbent paper

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Human CD117 molecule (CD117) ELISA kit

Procedure

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1. Preparation: Remove the kit from the refrigerator and let it reach room temperature for 30 minutes.

2. Dilute the 20× washing solution with distilled water to the working concentration.

3. Add standards and samples: Place the enzyme-labeled plate on the frame. Set up standard wells, sample wells, and blank control wells. Add 50μL of standard to each standard well, 10μL of sample and 40μL of diluent to each sample well (sample diluted 5 times), and leave blank wells empty.

4. Incubate at 37°C for 30 minutes.

5. Wash the plate: Discard liquid, blot dry, fill each well with washing solution, wait 1 minute, discard, repeat 4 times.

6. Add enzyme working solution (50μL per well), no addition to blank wells.

7. Incubate again for 30 minutes at 37°C.

8. Wash the plate again following step 5.

9. Add color reagent A (50μL per well), then reagent B (50μL). Incubate in the dark at 37°C for 15 minutes.

10. Stop the reaction by adding 50μL stop solution to each well. The color will change from blue to yellow.

11. Measure OD values at 450nm within 15 minutes after stopping the reaction.

12. Calculate: Use the standard curve to determine the sample concentration based on its OD value. Multiply by the dilution factor to get the actual concentration.

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Human CD117 molecule (CD117) ELISA kit

Sample requirements

1. Samples must not contain sodium azide (NaN₃), as it inhibits horseradish peroxidase (HRP).

2. Samples should be processed immediately after collection. If not tested right away, store at -20°C, avoiding repeated freezing and thawing.

3. Ensure samples are fully centrifuged, without hemolysis or particles.

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Human CD117 molecule (CD117) ELISA kit

Precautions

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1. Follow the manual strictly. Results must be based on microplate reader readings.

2. If the enzyme-labeled plate is not used up, store it in a sealed bag with desiccant.

3. It is recommended to perform double testing for all standards, samples, and blanks, taking an average to reduce error.

4. Remember that the sample was diluted 5 times; multiply the result by 5 to get the actual concentration.

5. The detection range is 0.1–200ng/ml. If out of range, extend the standard curve or dilute using a special diluent.

6. If the color is too light, increase the substrate incubation time slightly.

7. Avoid cross-contamination by using separate tips for standards, samples, and blanks. Do not reuse any reagents or sealing films.

8. Do not mix reagents from different batches.

9. Substrate B is sensitive to light—keep it protected from exposure.

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Human CD117 molecule (CD117) ELISA kit

Summary of procedures

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Prepare reagents, samples, and standards

Add samples and standards, incubate at 37°C for 30 minutes

Wash plate 4 times, add enzyme reagent, incubate 30 minutes

Wash again, add color reagents A and B, incubate 15 minutes

Add stop solution, read OD values within 15 minutes

Calculate results based on standard curve

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Human CD117 molecule (CD117) ELISA kit

Detection range

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0.1–200ng/ml

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Human CD117 molecule (CD117) ELISA kit

Specification

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96T/box, 48T/box

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Human CD117 molecule (CD117) ELISA kit

Storage

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Store at 2–8°C, away from light and moisture.

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Human CD117 molecule (CD117) ELISA kit

Shelf life

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6 months