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Description
Human FcεRIα 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 the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 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 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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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| 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 a capture antibody against high affinity IgE receptor a chain (FcεRIα). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of high affinity IgE receptor a chain (FcεRIα) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human High affinity IgE receptor a chain ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The high-affinity IgE receptor α chain (FcεRIα) is a protein encoded by the FCER1A gene. The high-affinity IgE receptor plays a central role in allergic diseases, linking allergens to mast cells and initiating the inflammatory and immediate hypersensitivity reactions that characterize conditions such as hay fever and asthma. An allergic reaction occurs when two or more IgE receptors are cross-linked by IgE molecules, which in turn bind to allergen (antigen) molecules. This perturbation releases histamine and proteases from the mast cell cytoplasm and leads to the synthesis of prostaglandins and leukotrienes—potent effectors of hypersensitivity reactions. The IgE receptor is composed of three subunits: α (the original protein), β, and γ. Only the α subunit is glycosylated. | |||||||||||||||||||||||||||||||||
| 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, tissue homogenates and other biological fluids |
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4.4 ★★★★★
Based on 23 reviews
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Product Reviews
★★★★★ 5
Better than you imagined...
I bought a 4'x6' from Safavieh 10 years ago and just received its replacement. I've slept on it every night and meditated on it every day. In short, I love it. It's the most comfortable surface I can imagine.
Here's what you need to know. A sheepskin is just that, animal pelt(s). The 4x6 is four pelts sewn together. It will have a sheepskin smell, which will reduce over time. When it comes, it is rolled up and in a heavy clear poly sleeve, Open the end, not the side! This way if you need to return it, you can roll and slide it right back into the shipping sleeve.
The dimensions you buy are approximate. It's a naturally variable product.
You might prefer a medical sheepskin if you live in a hot climate. The fur on these is altered to make the hairs shorter.
I think I will cover it with a bottom sheet from now on. It might last 20 years that way.
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Reviewed in the United States on October 13, 2024
★★★★★ 1
PACKAGE STUNK UP BLDG HALLWAY
I went downstairs to pick up the package from our building management office. I could smell it already. The rug was rolled up in clear plastic with one end exposed/open. From what I could touch, the rug felt soft and the color was good. But I was embarrassed to have received this and then bring it into the elevator with me. The smell was horrendous, like sewage or fecal matter.
BACKSTORY: I had ordered 2 different sheepskin rugs from 2 different brands. My first time buying sheepskin. The other has a very faint animal/hide smell that is detected only when right up against it. This Safavieh rug, however, was for a different location in the house. Even without the first rug for comparison, the smell of this one is unbearable. I've bought many wool and polypropylene rugs from Safavieh. So I did not expect a negative experience.
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Reviewed in the United States on September 25, 2020
★★★★★ 5
Great product: thick and cozy
Size: Single Pelt/2 ft x 3 ft, Color: Natural (Ivory Whiite)
This is a lovely, thick sheepskin. I’m using it to drape over a chair. It’s very soft. The color is a natural white. Arrived quickly. The price was great for such a wonderful sheepskin.
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Reviewed in the United States on April 3, 2026
★★★★★ 5
Love it!
Size: 2' x 6' (Double Pelt), Color: Linen, Size: 2' x 6' (Double Pelt), Color: Linen
Exceeded all expectations. Bigger than I imagined it would be, super soft and thick, beautiful color (Linen), and no funky smell at all. It seems very durable. After running my hands all over it, no shedding occurred. I have yet to sit on it myself, though, as my cat claimed it right away - didn't even wait for me to remove the tag! He's been there for hours and I'm thinking I've lost my snuggle buddy, so I bought two more in the smaller size so he can have his own on the bed with me at night. I am so amazed by the quality, feel, and color. I can't believe I almost let other reviews deter me from this purchase. 100% thrilled with this beautiful pelt. Absolutely recommend!
UPDATE:
I received the other 2 purchased. Just as soft, just as thick, consistent color (Linen), no shedding, no smell. They have been in use for several days now without any issues. Love them all - and my snuggle buddy is back! 🥰
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Reviewed in the United States on April 22, 2024
★★★★★ 5
It’s wonderful!!
Size: 2' x 3' (Single Pelt), Color: Brown, Size: 2' x 3' (Single Pelt), Color: Brown
The product is wonderful! You know it is real, coz the smell doesn’t go away. I use it to meditate and cannot how nicely it created the perfect comforting barrier to help elevate my experience. I however want to find alternatives coz I stopped using animal products after turning a vegetarian. The size (it is bigger in person than in the pic) and color(brown) are perfect for what I do and doesn’t need much/any maintenance!
The white however needs a lot of attention and care.I’m sure it would last a decade and the weight is just right too!
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Reviewed in the United States on April 16, 2026