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Description
Human IgG4 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect specimens using EDTA or heparin as an anticoagulant and centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-Test Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Sensitivity | 0.071 ng/mL | |||||||||||||||||||||||||||||||||
| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with an immunoglobulin G4 (IgG4) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. 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 immunoglobulin G4 (IgG4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Immunoglobulin G4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Immunoglobulin G4 (IgG4) is a relatively rare IgG subtype. IgG4 is produced by Th2 cells, which regulate B cells through the secretion of cytokines such as IL-4 and IL-13. It has low affinity for target antigens, cannot bind serum complement C1q, and has low affinity for Fc receptors. It cannot activate the classical complement pathway or cytokines, but it can inhibit the formation of immune complexes of other subtypes. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, cell culture supernatant |
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4.9 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 5
Best pillow I've ever had
Size: Queen (Pack of 1)
I don't really write reviews but holy buckets, Batman. This pillow is legit. Back, neck, and shoulder pain is nothing new for me; I've had scoliosis since I was a kid and hyperflexibility in my neck since birth leading to frequent muscle spasms that have caused me to miss work.
Guys. This pillow. It is the most comfortable pillow I've ever had. Like, my neck doesn't hurt and I genuinely didn't remember what that felt like. It's simultaneously supportive and soft and it doesn't sink in. The instructions are super clear to understand and the pillow fluffs up no problem if it gets a little flat. This is a game changer for me and I so hope anyone who buys this has the same experience.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 6, 2026
★★★★★ 5
I highly recommend this pillow, it’s great for someone with neck or back problems.
Size: Queen (Pack of 1), Size: Queen (Pack of 1)
I absolutely love this pillow! When it says it’s adjustable it definitely means that. When you first get it, it’s hard to believe that it’s going to be anything but put it in the dryer for a few minutes and it becomes so large. You will definitely have to remove some of the stuffing unless you like a really large fluffy pillow.
The stuffing is high-quality and once it has been put into the dryer it becomes a fibrous type material that can be fluffed over and over and go back to the same size. It seems to be a high-quality cover on the outer portion and is thick and has a slight cooling affect. It has a separate cover on the inner portion covering the foam stuffing.
I ordered this because I have neck problems and it has been super comfortable for me because I’m able to adjust it multiple times depending on my needs for the particular night when I lay down. Having the extra stuffing means that if the pillow does happen to go flat I can always add the extra that I took out when I received it.
It did have a slight odor when it first came, but after putting it in the dryer, it went away immediately. I do think that’s only due to the plastic that it’s shipped in not anything with the pillow.
I think this pillow is well worth it price and I would highly recommend it for anyone that is a side or back sleeper or has any type of neck problems.
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Reviewed in the United States on February 3, 2026
★★★★★ 5
Amazing!
Size: King (Pack of 1)
Such a great pillow I’m buying another one! You can add or subtract stuffing to your liking. It’s really nice quality, quilted and soft. Best pillow ever. Super comfy and great sleep. Pretty firm with all the stuffing. Perfect the way it came with stuffing on the side. My husband loves it and now I want one.
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Reviewed in the United States on May 15, 2026
★★★★★ 5
Totally worth the money
Size: King (Pack of 1), Size: King (Pack of 1)
Right off the bat, the packaging is absolutely adorable and felt like I was buying a premium product. I love that this pillow is adjustable and as a side sleeper, I needed a pillow that is both firm and supportive of my neck. This hits all the boxes. We will be buying more of these for our bed!
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Reviewed in the United States on June 1, 2026
★★★★★ 5
Deserves more Stars
Size: Queen
I was very hesitant to make a pillow purchase off Amazon. Turns out it’s the very best purchase! I had chronic neck issues the chiropractor couldn’t resolve. It was the first time, I had an unsolvable issue. I finally decided to spend the money and buy new pillows. My neck issue was 95% resolved the next morning. I’m not kidding! It’s now 100% resolved after a very short period of time. They are a wonderful gift to myself.! quality is top-notch. The support is fantastic. They adjustability is phenomenal and you can adjust the firmness because they give you extra foam. They’re completely worth every penny. I’m 61 years old, 100 pounds and very active. Waking up with neck strain was horrible.
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Reviewed in the United States on June 4, 2026
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