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Description
Human RBP3 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 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 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 Retinol Binding Protein 3, Interstitial (RBP3). 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 Retinol Binding Protein 3, Interstitial (RBP3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Retinol Binding Protein 3, Interstitial ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Retinol-binding protein 3 (RBP3), also known as IRBP, is a protein encoded by the RBP3 gene. It is specifically expressed by photoreceptors and is the most abundant protein in the interphotoreceptor matrix. It appears to play a role in the transport of retinoids during the visual cycle between the photoreceptor outer segments and the retinal pigment epithelium, and possibly between Müllerian cells. During transport, IRBP protects retinol from isomerization and oxidation. In addition to its role in the visual cycle, IRBP also binds fatty acids, suggesting its involvement in retinal development. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.3 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Daughter ranks it 5/5. I rank them 4/5 due to heel comfort.
Size: 8, Color: Black, Size: 8, Color: Black
My foot is 9.44 inches long which usually places me in a 7.5. For this chart, it suggested the size 8. It's a perfect fit. The back zipper is easy enough to use. You don't even really need to undo the top strap once you have it adjusted to your preferred spot. My foot is narrow to normal around the heel and I'm wider in the toe box. These heels accommodate that well. I have plenty of room around my toes. The toe strap isn't smashing down either. The shoes seems well-made. The straps do not have a threads hanging. There's no sloppy glue drips around the bed. The bottom is strong and the heel is solid. I'm not a huge fan of thicker/chunky heel looks usually. I like the skinner heel styles mostly but ... there's a big positive to the chunky heels. They're very easy to walk in. Great heels for non-formal occasions. The bottom is not slippery. The cushioning around the toes in very nice. My only complaint is that the cushioning by the heel feels inadequate- especially at this price. It feels like you're walking on a block of wood. For that reason, I gave them to my middle schooler. She's thrilled. She pranced around the house in them testing them out for herself. She adjusted well to walking in them and she finds them comfy.
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Reviewed in the United States on January 21, 2026
★★★★★ 5
Lucky Brand womens Herrika
Size: 9, Color: Smoke Grey
These shoes are some of the comfiest heels I've ever worn! The heel actually has a nice height to it, but it's at a gentle slope that feels very comfortable. The heel itself is thicker, which makes it easier to walk, and I love the ankle strap for added security. The shoe's construction is solid, the materials are durable, and the zipper at the back of the ankle moves smoothly. I got the smoke grey color, which I think is really more of a lighter skin tone color, so I don't really agree with the name, but the color is accurately represented by the photos on the product page. This is a great neutral shoe that will pair with many different outfits. It's high quality and versatile, so I think it's well worth the cost. I love my new shoes!
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Reviewed in the United States on January 15, 2026
★★★★★ 5
Well made, true to size, comfortable, and attractive looking!
Size: 12, Color: Black
Wife was wanting a new pair of heels for a work party she has coming up and I happened to see these and thought they strongly resembled what she was looking for so I got them as a surprise for her. They arrived a few days ago and she tried them on that night and wore them around the house and to work a couple times to see how she likes them. So far, she is extremely pleased with them! She said they fit perfect and she's had no issues with comfort wearing them all day for work. She really likes the wider heel as it gives her more stability. Her favorite thing so far though has been all the compliments/comments she's getting from the other ladies in her office about them.
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Reviewed in the United States on January 15, 2026
★★★★★ 5
Secure and super comfortable.
Size: 9, Color: Black, Size: 9, Color: Black
Lucky Brand nailed it with these sandals. The craftsmanship of the leather is top notch. I love the look of the crossover straps, they add a chic touch to any outfit. The fit is fantastic, I ordered my usual size 9 and found them to be perfectly true to size.
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Reviewed in the United States on January 16, 2026
★★★★★ 5
Get them you won’t be disappointed
Omg I am going to buy more of this shoe just to always have it before there gone and or unavailable! They are so comfy and stylish with any pair of jeans or a skit that I could wear!! Totally satisfied beyond words haha! 8.5 is my size and it fits perfectly! So happy I found this and took a chance Soda shoes never disappoint. One happy customer.
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Reviewed in the United States on March 8, 2026
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