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Description
Human GPRC5A ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 utilizes 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 retinoic acid-induced protein 3 (GPRC5A). 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 retinoic acid-induced protein 3 (GPRC5A) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Retinoic acid-induced protein 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | G protein-coupled receptor family C5 group member A (GPRC5A), also known as retinoic acid-induced protein 3 (RAI3), is a protein encoded by the GPRC5A gene. This gene and the mRNA it encodes were originally identified as a phorbol ester-induced gene and named phorbol ester-induced gene 1 (PEIG1). Two years later, it was rediscovered as a retinoic acid-induced gene and named retinoic acid-induced gene 1 (RAIG1). The protein it encodes was later named retinoic acid-induced protein 3. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.2 ★★★★★
Based on 49 reviews
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Product Reviews
★★★★★ 3
Very comfortable wonderful chair
I like the chair it was very easy to assemble makes no noise when I'm going across the floor the size was perfect for me the stability was great the screw quality was very easy it's all very well put together got it on sale value for the money very good how do you just meant no problems it was great
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Reviewed in the United States on September 14, 2024
★★★★★ 5
excellent quality, Good Support
I got this on sale for $67. The process to assemble it was very straightforward and used an Allen wrench. It was rated as the best item in its category and had five stars. I wouldn't consider it a gaming chair, Although it tries to be one. It feels more like an office chair that was glam up to be a gaming chair. But it does an excellent job at supporting a tall large individual, Excellent Support, Good posture. Worth every penny, But not in the same category as a typical gaming chair. Lack some of the bells and whistles had a typical gaming chair would have, But every bit the money I paid for it. If you want something nicer then you need to buy like a $200+ Gaming chair with the bells and whistles.
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Reviewed in the United States on June 13, 2024
★★★★★ 5
Absolutely LOVEEE
I loooove this chair. I work from home and it is so comfortable to sit in any position. It is great for sitting criss cross but you can also sit normal or on your knees. Very stable and easy to put together. Great quality
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Reviewed in the United States on April 21, 2026
★★★★★ 5
I LOVE MY Criss Cross Applesauce Office Desk Chair IS THE BEST
Color: Cream, Size: PU Leather(No Wheels)
The Criss Cross Applesauce Office Desk Chair is a unique and innovative addition to any workspace, offering a blend of comfort, functionality, and modern design.
What immediately stands out about this chair is its ergonomic design. The criss-cross frame provides excellent support to the back and promotes proper posture during long hours of sitting. This feature is particularly beneficial for reducing strain on the lower back and promoting overall spinal health, which is essential for those who spend extended periods at their desks.
The chair is upholstered with high-quality materials that offer both durability and comfort. The cushioned seat and backrest provide ample support while maintaining a plush feel, ensuring comfort throughout the workday.
Another notable feature is the adjustable height mechanism, which allows users to customize the chair to their preferred seating position. This versatility ensures that the chair can accommodate individuals of varying heights and preferences, enhancing overall ergonomic comfort.
In terms of aesthetics, the Criss Cross Applesauce chair boasts a modern and sleek design that complements contemporary office decor. Its minimalist appearance blends seamlessly into any workspace environment, adding a touch of style without compromising functionality.
Assembly of the chair is straightforward, with clear instructions provided for easy setup. The sturdy construction and stable base ensure reliability and safety, providing peace of mind for users.
While the chair excels in comfort and design, some users may find the criss-cross frame design to be unconventional compared to traditional office chairs. It's important to consider personal preferences and ergonomic needs when choosing a chair that best suits individual comfort requirements.
Overall, the Criss Cross Applesauce Office Desk Chair combines ergonomic support with modern aesthetics, making it a practical and stylish choice for enhancing comfort and productivity in the workplace. Its blend of functionality, comfort, and contemporary design earns it a recommendation for anyone looking to upgrade their office seating experience.
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Reviewed in the United States on July 10, 2024
★★★★★ 5
Perfect little black chair
I bought this chair for four reasons:
1. My favorite color is black, but the company offers different color options.
2. The height of the chair was exactly what I was looking for. I did not want a tall back for the chair. The chair is close to the ground, which is a plus for me, my back, and my knees as I get older.
3. The chair comes with the option of wheels or no wheels; I chose the no wheels option, as I get dizzy and sick when I spin around for too long.
4. The chair’s wide back and seat design are perfect for me, since I prefer to sit criss-cross in a chair.
Pluses: It reclines, is comfy, and can be cleaned and moved easily.
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Reviewed in the United States on January 7, 2026