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Description
Rat apo-D ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 3. 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 200 ng/mL). Then dilute to the following concentrations: 200 ng/mL, 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.125 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 200ng/mL standard working solution into the first EP tube and mix thoroughly to make a 100ng/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 competitive enzyme-linked immunosorbent assay (ELISA). Samples, standards, biotin-labeled antibodies, and HRP enzyme conjugates are added sequentially to microwells pre-coated with universal species Apolipoprotein D (apo-D) antigens. After incubation and washing, the substrate TMB is used for color development. TMB is converted to blue under the catalysis of peroxidase (HRP) and to the final yellow under the action of acid. The depth of the color is negatively correlated with the universal species Apolipoprotein D (apo-D) in the sample. The absorbance (OD value) is measured at a wavelength of 450nm using an enzyme reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Apolipoprotein D ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Apolipoprotein D (apo-D) is a 25 to 30 kDa glycosylated protein and a member of the lipid protein superfamily. As a transporter of several small, hydrophobic molecules, its known biological functions are primarily related to lipid metabolism and neuroprotection. It is a multiligand, multifunctional protein involved in lipid trafficking, food intake, inflammation, antioxidant responses, and the development of various cancer types. A key aspect of its role in lipid metabolism involves the transport of arachidonic acid and the regulation of eicosanoid production and delivery in metabolic tissues. Apo-D expression in metabolic tissues correlates positively and negatively with insulin sensitivity and glucose homeostasis in a tissue-dependent manner. | |||||||||||||||||||||||||||||||||
| 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 | 3.12-200ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.1 ★★★★★
Based on 1818 reviews
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Product Reviews
★★★★★ 5
That ending 😫
Format: Kindle
I fell into a false sense of security and really thought this was gearing towards a happy ending. Then I realized there’s no work they don’t punish Andrew. I really liked Vale’s character. I don’t normally read books with pregnancy but going into this knowing she was pregnant made it more enjoyable for me.
I loved Bishops devotion to her and her happiness. I also loved that Holt and Mercy couldn’t fight their attraction to her. I love scent matches so very much. I’m so curious to see how this duet will end up. And I need to pay more attention and notice that a book I’m starting is a duet to begin with lol
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 21, 2025
★★★★★ 5
oh wow
Format: Kindle
I just knew there was something about Cooper! I’m wondering if he’s about to be included but damn I’m glad he’s at least not a rapist and creepy guy, he just got called on assignment and had to go! This should be interesting! She’s gonna run and then what’s his face is gonna grab her. I’m worried! Wow that was a great book and cliffhanger! Loving this!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 27, 2025
★★★★★ 5
A good read
Format: Kindle
A good read, just fluffy cuteness, no antagonism. I like all the characters. It could have used another round of editing however, glanfds being one error that cracked me up, and my personal pet peeve was that the author kept using the word fill instead of feel, which I promise you are not interchangeable haha, but it's definitely better than the majority of books I read on here mistake-wise.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 28, 2024
★★★★★ 4
amazing
Format: Kindle
Knot the Bride was a fantastic read! The characters were all amazing and well-developed. It was easy to like them all. Sophia, Luca, Nick, and Gavin were all perfect for each other. It was such a charming story that had me hooked the entire time. I did wish there were POVs from Luca, Nick, and Gavin but it was still an amazing book without it.
I am excited to read the next book in the Willowside Omegaverse series! This is definitely a must-read for fans of omegaverse romance!.
I received a free copy of this book and am voluntarily leaving a review.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 25, 2025
★★★★★ 3
3 Star Read,
Format: Kindle
This book wasn't bad, but wasn't my cup of tea. It's highly disappointing because the storyline is so original. There is no real conflict or resolution, so the entire thing feels flat.
As a lover of omegaverse books, I know there is a ton of variety out there, and ov is really up to the author. But this one is weird. Omegas have multiple scent glands all over their bodies and go into week long heats every month. Alphas have knots in the middle of their shaft instead of the base, and the knot doesn't always swell, no explanation of when or why. It doesn't engage at all when the mouth is in play.
I also didn't enjoy the author's writing style. Each paragraph is only 1 or 2 sentences long, and the entire book reads very stacato. The conversations are stiff and unnatural feeling. Everything is very repetitive, both in word choice and in thought. The same thing is repeated 3 or 4 times over a single page, multiple times over. I ended up doing so much skimming.
The first 50% of the book is all slow burn, and the last 50% is almost straight mediocre spice. This wouldn't have been all bad if the grammar and spelling errors didn't start at the exact same time. Tongue is repeatedly misspelled in the middle of the spice.
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Reviewed in the United States on September 14, 2024
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