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Description
Mouse SPR 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 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 Sepiapterin Reductase (SPR). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Sepiapterin Reductase (SPR) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Sepiapterin Reductase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Sepipterin reductase, also known as SPR, is an enzyme encoded by the SPR gene. It catalyzes the NADPH-dependent reduction of various carbonyls, including pteridine derivatives, and belongs to a group of enzymes known as aldo-keto reductases. It plays an important role in the biosynthesis of tetrahydrobiopterin. Sepipterin reductase deficiency is a rare disease. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.1 ★★★★★
Based on 1630 reviews
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Product Reviews
★★★★★ 5
Transformative for Dry, Cracked Heels!
Color: Original Grey
I recently purchased Dr. Frederick's Original Moisturizing Heel Socks, and they have truly transformed the condition of my feet! I can’t recommend them highly enough.
Effective Moisturization: These socks are infused with moisturizing agents that work wonders. After just a few uses, I noticed a significant reduction in dryness and cracking on my heels. My feet feel softer and smoother, and I love that I can wake up to beautifully moisturized skin.
Comfortable Fit: The socks are incredibly comfortable to wear, with a snug yet gentle fit. I appreciate that they don’t feel restrictive, allowing me to walk around my home while wearing them without any discomfort. They are also lightweight, making them easy to slip on before bed.
User-Friendly: They are super easy to use—just wear them overnight, and let the socks do their magic. I love the simplicity of incorporating them into my nighttime routine.
Durable Quality: The quality of the socks is impressive. They wash well and have maintained their shape and effectiveness after multiple uses. I feel confident that they will last a long time.
Final Thoughts: If you're struggling with dry, cracked heels, I highly recommend Dr. Frederick's Original Moisturizing Heel Socks. They are a fantastic investment in foot care and have made a noticeable difference in the health of my skin. These socks have become a must-have in my self-care routine!
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Reviewed in the United States on October 4, 2024
★★★★★ 5
These worked for me!
Color: Original Grey
They worked great. I bought these for cracked sore heels. It took 2-3 weeks but my heels no longer hurt and the cracks have closed and look more like an indention than a crack now. They feel so much better, but I'll probably wear them another week just to be sure they stay healed. They look and feel like regular socks from the outside but only cover the heel with the toe open. Not think but not thick, again like regular socks. They would probably be slippery on a smooth surface the same as socks. The inside feels like rubber or something smooth at the heel. I have washed them once each pair and accidentally dried one pair in the dryer. They still seem to feel the same so I think they're fine. They're pretty inexpensive and had different colors and sizes.
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Reviewed in the United States on February 10, 2026
★★★★★ 4
Truly Helped My Cracked Heels
Color: Original Grey
I didn't include a photo cause they look exactly like their advertisement photo.
I so liked the silicon-type heel lining as it kept the heel-repair cream on my heel and did not get soaked up by the sock. But, it is so, so slippery when walking around in them! Take care to be mindful when you set each foot down as the lotion can slide your heel across the sock quite a bit. If these socks were for an elderly person, I would have them only sit while using the sock - no walking around for fear of falling.
I have larger ankles and found that having both the top and the bottom hems/elastic, etc. the same size to be quite troublesome and was not a nice feeling or fit on my upper hem that hit my ankle. Don't get me wrong, I am so glad that I got them and I continue to use them and they are helping my horribly-cracked heels greatly, but I find that I have to keep moving the top hem up and down so that it doesn't stay in one position as it cuts in to my ankle.
My feedback and advice might be to please make the upper hem somewhat larger than the one that is on the bottom and goes over my actual foot. They really should not be the same circumference...or else make different sizes - S, M, L, etc. That's really my only complaint or "con" to ordering these.
I think they are a great concept and I like the fact that I can throw them in the washer and dryer.
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Reviewed in the United States on June 8, 2024
★★★★★ 5
Actually works, and really well
Color: Original Grey
I bought these because I got them on a great deal and was quite skeptical, but they actually worked exactly as advertised and I saw a huge difference after the first night! I understand why they don’t cover the whole foot like a regular sock (it would be stifling), but it would be incredible if they made a pair that go forward enough to cover the ball of the foot. One pair lasted about 3 weeks before the moisturizer wore out (I did notice a dropoff after washing, so for best results put them on when you’re clean and just for overnight, so you can avoid washing).
Apologies for no photos- no one wants to see the before photos. These work so well I’ve just come back to order more even though I’m now paying full price. Definitely, definitely recommend.
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Reviewed in the United States on May 28, 2026
★★★★★ 5
Comfortably and works
Color: Original Grey
❤️ I used these with magnesium lotion on my heals for 3 nights, soaked my feet every morning and cracked heals are gone!
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Reviewed in the United States on April 25, 2026