Human Anti-PEG IgG ELISA,specification,price,image-Bio-Equip in China

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Product Specification

Usage

Specimen Requirements

1. Specimens should be extracted as soon as possible after collection, following the relevant literature. Experiments should be performed as soon as possible after extraction. If experiments cannot be performed immediately, specimens can be stored at -20°C, but repeated freezing and thawing should be avoided.

2. Samples containing NaN3 cannot be tested, as NaN3 inhibits horseradish peroxidase (HRP) activity.

Procedure

1. Dilution of Standard: This kit provides one standard at full concentration. Users can dilute the standard in a small test tube according to the chart below.

2. Sample Addition: Set up a blank well (no sample or enzyme-labeled reagent is added to the blank control well; all other steps remain the same), a standard well, and a well for the sample to be tested. Accurately add 50 μl of the standard sample to the ELISA-coated plate. First, add 40 μl of the sample diluent to the sample wells to be tested, followed by 10 μl of the sample to be tested (final sample dilution is 5-fold). Add the sample to the bottom of the plate well, avoiding contact with the well walls. Gently shake to mix.

3. Incubation: Seal the plate with a sealing film and incubate at 37°C for 30 minutes.

4. Prepare the solution: Dilute the 30x concentrated wash solution 30x with distilled water and set aside.

5. Wash: Carefully remove the sealing film, discard the liquid, and spin dry. Fill each well with wash solution. Let it sit for 30 seconds before discarding. Repeat this process 5 times and pat dry. 6. Enzyme Addition: Add 50 μl of enzyme-labeled reagent to each well, except for the blank well. 7. Incubation: Same as in step 3. 8. Washing: Same as in step 5. 9. Color Development: Add 50 μl of Color Developing Reagent A to each well, followed by 50 μl of Color Developing Reagent B. Gently shake to mix. Incubate at 37°C in the dark for 10 minutes. 10. Stop: Add 50 μl of Stop Solution to each well to terminate the reaction (the blue color will immediately turn yellow). 11. Measurement: Use the blank well as the zero setting and measure the absorbance (OD) of each well at 450 nm. Measurement should be performed within 15 minutes after adding the Stop Solution. Calculation: Draw a standard curve on graph paper with the standard concentration as the horizontal axis and the OD value as the vertical axis. Use the standard curve to find the corresponding concentration based on the sample's OD value; then multiply it by the dilution factor. Alternatively, use the standard concentration and OD value to calculate the linear regression equation for the standard curve. Substitute the sample's OD value into the equation to calculate the sample concentration, and then multiply it by the dilution factor to obtain the actual sample concentration.

Sensitivity

10pg/ml

Species Reactivity

Human

Theory

This kit uses a double-antigen sandwich assay to measure the level of human anti-PEG IgG in samples. Purified antigen is coated on a microplate to create a solid-phase antigen. Anti-PEG IgG is then added sequentially to the coated wells. Anti-PEG IgG then binds to the HRP-labeled antigen, forming an antigen-antibody-enzyme-labeled antigen complex. After thorough washing, the plate is then treated with the substrate TMB for color development. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of anti-PEG IgG in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader, and the concentration of human anti-PEG IgG in the sample is calculated using a standard curve.

Detection Type

Used to determine the content of anti-polyethylene glycol immunoglobulin G (Anti-PEG IgG) in human serum, plasma and related liquid samples.

Composition

Serial number	Component name	Specification	Serial number	Component name	Specification
1	30 times concentrated detergent	20ml×1 bottle	6	Developer B solution	6ml×1/bottle
2	Enzyme Labeled Reagent	6ml×1 bottle	7	Stop Solution	6ml×1 bottle
3	Enzyme-coated plate	12 wells×8 strips	8	Standard (4000pg/ml)	0.5ml×1 bottle
4	Sample diluent	6ml×1 bottle	9	Standard diluent	1.5ml×1 bottle
5	Developer A Liquid	6ml×1 bottle	10	Sealing film	2 photos

General Notes

1. After removing the kit from the refrigerator, allow it to equilibrate at room temperature for 1 hour before use. If the enzyme-coated plate is opened but not completely used, store it in a sealed bag.
2. Crystals may form in the concentrated wash solution. Warming in a water bath during dilution can aid dissolution. This will not affect the results during washing.
3. Use a pipette for each sample addition step and frequently calibrate its accuracy to avoid experimental error. The time for each addition should ideally be within 5 minutes. For large numbers of samples, using a dispenser is recommended.
4. Develop a standard curve with each measurement, preferably in duplicate. If the analyte concentration in the sample is too high (the OD value of the sample is greater than the OD value of the first standard well), dilute the sample a certain number of times (n) with sample diluent before measurement. When calculating the total dilution factor (×n×5), multiply the final dilution factor.
5. Plate sealing film is for single use only to avoid cross-contamination.
6. Protect the substrate from light. 7. Strictly follow the instructions for use. Test results must be based on the readings of the microplate reader. 8. All samples, wash solutions, and waste should be treated as infectious agents. 9. Components from different batches of this reagent must not be mixed.

Storage Temp.

Unopened test kit, stored at 2-8°C, has a shelf life of 6 months.

Test Range

40pg/ml –2200pg/ml