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Description
Mouse CD5L 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: 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: 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. 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. 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 CD5 Antigen-Like Protein (CD5L) capture antibody. 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 CD5 Antigen-Like Protein (CD5L) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse CD5 Antigen Like Protein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | CD5L is a key regulator of lipid synthesis. It is primarily expressed by macrophages in lymphoid and inflammatory tissues, modulating inflammatory responses such as infection or atherosclerosis. It also inhibits lipid droplet size in adipocytes. After entering mature adipocytes via CD36-mediated endocytosis, it binds to the cell membrane enzyme FASN, inhibiting the activity of fatty acid synthase, leading to lipolysis and the degradation of triglycerides into glycerol and free fatty acids (FFA). CD5L-induced lipolysis occurs with the progression of obesity. Following the recruitment of inflammatory macrophages to adipose tissue, CD5L contributes to obesity-related inflammation, which contributes to insulin resistance and obesity-related metabolic diseases. CD5L-mediated intracellular lipid regulation has a direct impact on transcriptional regulation mediated by the nuclear receptor ROR-gamma (RORC). | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, and other biological fluids |
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4.3 ★★★★★
Based on 8 reviews
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Product Reviews
★★★★★ 4
A Thoughtful and Compelling Romance with Depth and Maturity
Format: Kindle
Miranda MacLeod and TB Markinson deliver a compelling and emotionally charged romance in The Anatomy of Forever, a novel that successfully balances personal transformation, ethical dilemmas, and the weight of professional responsibility.
Dr. Rowan Colchester, newly embracing her true self after a difficult divorce, is not prepared for the whirlwind that follows a passionate encounter with a younger woman. When fate reunites her with Dr. Simone Doucette, an idealistic doctor seeking solace in a Cape Cod ER, their undeniable connection becomes fraught with complications. The revelation of their unexpected personal link adds a layer of tension that challenges not only their burgeoning relationship but also their careers and reputations. As the stakes rise, Rowan and Simone must navigate professional ethics, small-town scrutiny, and their own vulnerabilities to fight for a second chance at love.
While the “sleeping with your daughter’s best friend” trope initially presents a hurdle, the novel compensates with a well-developed plot that offers depth beyond the romance. MacLeod and Markinson craft a narrative rich with stakes that extend beyond the personal, grounding the story in a high-stakes professional and community-driven context. The characters are well-drawn and compelling, both Rowan and Simone stand out as confident, determined, and capable women who refuse to be defined by societal expectations or past mistakes. Even secondary characters, such as Erica, evolve in ways that add nuance to the story.
A refreshing aspect of The Anatomy of Forever is its portrayal of strong, independent women who communicate effectively and fight for what they want. Rather than relying on contrived misunderstandings or overplayed angst, the novel explores mature relationship dynamics, making the romance feel authentic and rewarding.
Overall, this is a well-executed sapphic romance that offers substance, well-rounded characters, and a satisfying blend of personal and professional stakes. MacLeod and Markinson have crafted a narrative that is engaging, thought-provoking, and ultimately, deeply satisfying.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 25, 2025
★★★★★ 5
Lives up to all the hype
Format: Kindle
Stars: 5 Stars
Format: Audiobook
Narrator: Lori Prince
Series: Stand Alone
Steam Level: Steamy
I read this for the 2026 A to Z Book Challenge for A.
This was the first book I added to the A to Z Challenge because I was so excited to read it and it absolutely lived up to all the hype.
Quick Thoughts:
- There is something about the “we had a one night stand but didn’t know we both have the same person who is important to us in our personal lives” set up that I just have so much fun reading
- The trauma that Simone went through as a kid then again as an adult was absolutely heartbreaking
- Two things I really liked about this book is that it didn’t end when they got together and that the Big Third Act Drama wasn’t about them being together
- This is exactly how cats get adopted
- No third act break up!
- I hope Lance ends up in prison
- The character growth almost everyone in the book goes through in this was wonderful to watch
- I had the biggest grin with this ending. It was sweet and adorable and perfect and just made me so happy
- It is ALWAYS a joy to listen to Lori Prince. She did a fantastic job, as usual, and I sincerely hope to listen to a thousand more books of hers
Overall I loved this book. T.B. Markinson and
Miranda MacLeod, by themselves, are an auto buy but when they write together I actively stalk their new releases to preorder it the second I can. I can honestly recommend any book of theirs without having read it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 14, 2026
★★★★★ 5
A Story Of A Two Conflicted Powerhouse Woman!
Format: Kindle
The writing dynamic duo has once again brought us a story so big in chemistry and family. Two very different woman meet in a bar and decide to have a night they will remember. Although there is intense chemistry, one is leaving for another country and the other is replacing her ex in his powerful position. Two years later, these two woman meet again but it is where they both work and one is best friends with the others step daughter and the other is the boss. A web neither wants to cross but the hearts have other ideas. The supporting cast to these two women are just as good as the lead. But each time they get closer to a maybe, something comes between them which brings more problems. A well worth read. It has laughs, tears and family drama. Another great score for the writing duo.
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Reviewed in the United States on April 1, 2025
★★★★★ 5
The dynamic duo has done it again!
Format: Kindle
I think TB & Miranda are my favorite writing duo. I love all their books whether written independently or collaboratively. Although this is an age-gap story of two doctors, Rowan and Simone, it's a mature telling of two very independent yet professional women with a lot at stake in their careers, as well as their personal lives.
In addition to the MCs, the secondary characters added extremely juicy conflict and delightful humor and levity. I especially loved Marigold and her free spirited young daughter Poppy.
I wasn't sure how the ethical dilemmas would be solved, but these fabulous authors wrote a very thought-provoking and satisfying ending. Well done, you two! I wholeheartedly recommend!!!
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Reviewed in the United States on April 1, 2025
★★★★★ 4
Kittens. crazy kid, what could go wrong?
Format: Kindle
Good read. Usually I eschew romance unless it comes with a mystery or intrigue. T.B. Miranda, thank you for a very enjoyable book.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 13, 2025
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