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Description
Rat CPR 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 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 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 Cytochrome P450 Reductase (CPR) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Cytochrome P450 Reductase (CPR) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Cytochrome P450 Reductase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cytochrome P450 oxidoreductase (CPR), also known as NADPH:hemoprotein oxidoreductase, NADPH:hemoprotein oxidoreductase, NADPH:P450 oxidoreductase, P450 reductase, POR, CPR, CYPOR, and others, is the enzyme encoded by the CPR gene. It is a membrane-bound enzyme required for electron transfer from NADPH to cytochrome P450 and other heme proteins, including heme oxygenase and cytochrome B5, in the endoplasmic reticulum of eukaryotic cells. It also reduces other proteins and molecules, including cytochrome C and ferrocyanide. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.5 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 5
Great product!
Style: Deep Moisturizing, Size: 1.76 Fl Oz (Pack of 1)
I love this stuff! My mom introduced this to me while I was visiting her, and within 3 days of using it I knew I needed to order one for myself. It has no smell, goes on buttery soft, and soaks in well. It is not heavy at all. At night I layer it on thicker and during the day I put a very thin layer on under my makeup. My skin is so much softer and hydrated. I have had small bumps on my cheeks for so long and they were gone within 3 days. My skin looks dewy and soft.
I read reviews of other brands that smelled, were oily, or too heavy. This is none of those things and it is a great price!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 25, 2026
★★★★★ 5
Best Beef Tallow Cream I’ve Tried
Style: Deep Moisturizing, Size: 1.76 Fl Oz (Pack of 1)
Exactly what i was looking for, some of the beef tallow creams have a weird consistency which i don’t love wearing during the day. This cream is perfect for day and night. smells very good, application is perfect not greasy, moisturizing, soft. I love it! 10/10,
will be buying again 💗
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Reviewed in the United States on May 29, 2026
★★★★★ 5
Order it! Its great
Style: Deep Moisturizing, Size: 1.76 Fl Oz (Pack of 1)
I love the texture! Its silky smooth and light on the skin, no weird smell, very moisturizing. Ive used one of the containers for most of my pregnancy to keep the skin on my belly and hips moisturized. I add a few drops of organic frankincense to a nickel-sized amount of this balm and mix it together in my hands before applying. I use a clean q-tip to scoop out the tallow balm as needed too to be sure the rest in the jar stays clean, rather than using my washed fingers, to reduce accidentally contaminating the rest of it somehow. My skin has stayed so healthy thanks to this product. It reduces the itchiness of my expanding skin from the baby growing in my belly, and i think its made a positive difference in preventing stretch marks by keeping my skin hydrated as i use it almost daily and after every shower. I just got my second jar in the mail to use for this last month of my pregnancy, and I will probably use it for my baby when hes born too.
I also love that it comes in nice packaging, with a thank you note, very minimal plastic, and classy label. I dont think the jar is plastic either which i LOVE (pretty sure its glass container?) Thank you to the creators of this product. I kind of wish i could get it in a glass dispenser like lotion but i really dont care at the end of the day. Its amazing. You should try it.
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Reviewed in the United States on May 31, 2026
★★★★★ 5
Smells nice and light
Style: Deep Moisturizing, Size: 1.76 Fl Oz (Pack of 1)
Nice and creamy. Feels silky. Unfortunately, it does not help my knuckles very much. But I would buy it again because it is a good cream.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 15, 2026
★★★★★ 5
Excelentt
Style: Deep Moisturizing, Size: 1.76 Fl Oz (Pack of 1), Style: Deep Moisturizing, Size: 1.76 Fl Oz (Pack of 1)
This product is really good, I researched it because it is the closest to human skin, and that was what it did help me repair my skin that was injured after an allergy of more than two months. I felt that I had lost the elasticity and shape of my skin, this fat stuck to me and made me feel safe again.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 24, 2026
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