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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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4.9 ★★★★★
Based on 2083 reviews
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Product Reviews
★★★★★ 5
Great quality tux for the price
Color: White, Size: XX-Large, Color: White, Size: XX-Large
I was pleasantly surprised with the quality and fit right out of the package. The cloth has a subtle pattern that gives it an upscale look without being flashy, and overall it feels far nicer than I expected at this price.
Jacket:
The jacket has a good weight and thickness, so it feels solid and well-made. Sleeve length was perfect right out of the box, which almost never happens for me. The only fit issue was a bit of snugness in the chest and underarm area, but I have broad shoulders and a larger chest, so that’s not unusual. My dress shirt sleeve length is 36.5", so I went with an XXL instead of my usual XL to make sure the sleeves were long enough. That was the right call, because it’s always easier for a tailor to size something down than to try and make something too small work. Aside from a quick tweak in the chest, this could easily be worn straight out of the package.
Pants:
The pants are noticeably thinner than the jacket and you can kind of see through them, so you’ll want to be mindful when choosing undergarments. That said, the lighter fabric is actually a plus for summer outdoor events or warm indoor venues since it will help keep you cool and comfortable. The flexible waistband made them very comfortable, and the fit in both the waist and length was spot on. For reference, I usually wear 38x32 dress pants and these matched perfectly.
Overall:
The quality is excellent for the cost. Even with a minor tailoring bill, my total cost to own this tux will still be less than renting one for a single event. I couldn’t be happier with the purchase and wouldn’t hesitate to pick up another.
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Reviewed in the United States on August 13, 2025
★★★★★ 5
brings me to today’s formal fashion
Color: Brown, Size: X-Large
the peak lapel made me feel sharp for the big family dinner. the slim fit worried me, but it was surprisingly comfy, not too tight on my retired belly. the pockets are deep, which is great for my reading glasses case. didn't need much ironing, just hung it up. a solid, non-fussy suit for us old guys.
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Reviewed in the United States on October 26, 2025
★★★★★ 4
Works for Dress-Up, Not for Formal Events
Color: Brown, Size: Medium, Color: Brown, Size: Medium
This brown tuxedo is about what you’d expect for under $100 on Amazon. The fabric has a pretty brocade pattern but it’s very thin and floppy. The jacket is lined, which helps give it some structure and makes it look a lot nicer, but the pants don’t lay well since they’re unlined and lack structure. The brown color is actually really nice—rich without looking too chocolately, almost leaning burgundy. That said, the fit on both the jacket and pants is surprisingly good. Overall, this is a fun tux for a costume party or one-off dress-up event, but not something you’d want to rely on for a wedding or a truly formal occasion. Just keep your expectations aligned with the price.
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Reviewed in the United States on September 14, 2025
★★★★★ 5
A true beauty
Color: Red, Size: Large
This tuxedo set is so beautiful. The red is extremely striking and is sure to grab everyone's attention. I ordered a size L, as I am on the cusp of Medium and Large. The pants fit very well, and they have some adjustment points around the waist in case they need to be let out a little more. Also, in the waist band is some elastic portions, which make finding the perfect fit something you won't have to think much about.--- they will stretch with you. The jacket feels like it is custom fitted by a tailor. The quality of this tuxedo, considering the price point, is exceptional.
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Reviewed in the United States on September 6, 2025
★★★★★ 5
Its great
Color: White, Size: Large
Got it for my brother and he digs it. Says its comfortable and not too confining. It looks great. Definitely worth the money.
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Reviewed in the United States on September 20, 2025