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Description
Human ANGPTL2 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 the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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 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 angiopoietin-like protein 2 (ANGPTL2) 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 angiopoietin-like protein 2 (ANGPTL2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Angiopoietin Like Protein 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Angiopoietin-like protein 2 (ANGPTL2), also known as angiopoietin-like protein 2, is a protein encoded by the ANGPTL2 gene in humans. Angiopoietin-like protein 2 maintains tissue homeostasis by promoting adaptive inflammation and subsequent tissue remodeling. Excessive activation of ANGPTL2 induced by chronic stress promotes the disruption of tissue homeostasis caused by chronic inflammation and the development of metabolic diseases. Angiopoietin is a member of the vascular endothelial growth factor family and the only known growth factor that primarily targets the vascular endothelium. Angiopoietin-1, angiopoietin-2, and angiopoietin-4 are involved in angiogenesis. The ANGPTL2 protein is a secreted glycoprotein that shares homology with angiopoietin and may exert its effects on endothelial cells through autocrine or paracrine effects. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.0 ★★★★★
Based on 1696 reviews
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Product Reviews
★★★★★ 4
I finally took the plunge. A satisfying star read
Format: Kindle
What a chicken I was to leave off reading this book for YEARS because I couldn’t cope with Logan’s “situation” (no spoilers!) & how it could possibly be sorted in his own book. Oh ye of little faith! Elle Kennedy isn’t a #1 author by CHANCE! Oh no, that is TALENT baby! She took the end of book one, ran with it, then flipped that mother sucker on its head to dig deeper into Logan and learn about Grace. These two were definitely made for each other.
The story spans from one school year to the next and we get lots of context too. There are some side characters that I’m not sure o know why they were added as they didn’t add to the story and a particular side character that really needed an ending as she just disappeared. Why was she so unhappy? What was she getting into off page? Was she ever forgiven?
The s story seemed to end abruptly, and that threw me off a bit. I kept going like I was gonna flip another page, and that was the end! Besides the fact of feeling like there are so many things left unsaid this was a good book. Definitely liked book one more but that could also be because the show just came out on prime but nahh, I loved to book one even a decade ago, and it’s still amazing to this day!
The hard part of this was the female narrator on the portions when I listened…Lorelei Avalon, she was NOT CAST properly in my opinion at all. She’s not a bad narrator, she’s just not suited, even remotely, for a late teens early 20-year-old college student. Her voice is more for characters late 40s early 50s in my opinion. I’m glad on book 3. It looks like we changed the narrator so I hope to mostly listen to that one. As always, Lee Samuels did a phenomenal job. He’s been in the game for a while, so maybe that’s what helps make it right for him. But he definitely knew how to change the nuances of his voice to suit whatever character he was reading at that time.
All in all, this was a really good book that I’m sad. I waited so long to read. Thanks to campus diaries coming out on TV? I’m ready to finally finish up this series! Book 3 here I come!!
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Reviewed in the United States on May 28, 2026
★★★★★ 5
🎧
Format: Paperback
The start of this book had me nervous but I ended up really loving this book! I loved how aware Logan was to what he was doing to Grace and stopped it even though it hurt to read. I loved that Grace made him WORK for her forgiveness and some parts were even SO funny. I loved all four of the guys and their great banter. My sweet baby Logan really was going through it in his personal life and his story resonated so much with me. I cried multiple times while reading and cried happy tears at the ending. It was perfect and exactly how I hoped it would end.
🎧 The narrators did a fantastic job with the dual narration.
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Reviewed in the United States on May 17, 2026
★★★★★ 4
*MILD SPOILERS*
Format: Kindle
this was amazing!!! i loved the characters and the writing is a lot better than most of its sport-romance-book counterparts. the worst part of it for me was the climax, i felt as though the FMC’s actions did not match her personality type. he is unsure about their relationship and is worried that she will leave him due to the seemingly bleak and boring future ahead of him. he voices this to her and she gets mad. mind you, this man had to grovel and beg and complete tasks for a single DATE. like girl, if we had been dating for 2 months and i had to do all that to get a date, i would be insecure too. it was quick so easily ignorable, but still bumped it down a star.
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Reviewed in the United States on May 20, 2026
★★★★★ 5
The 1st one is better than this one but I still love it
Format: Kindle
The Mistake is the second book in Elle Kennedy’s Off-Campus series, following a new couple while maintaining the humor, steamy romance, and emotional depth that made The Deal so popular. This installment focuses on Logan, Garrett’s best friend and fellow hockey player, and Grace, a quiet but strong-willed freshman, as they navigate second chances and personal growth.
John Logan – Logan is charming, funny, and a ladies' man, but beneath his confident exterior, he struggles with deep-seated family issues and a sense of being trapped by his circumstances. His development throughout the book makes him more than just the stereotypical player.
Grace Ivers – A freshman who starts off shy but proves to be independent and assertive. She isn’t easily won over, and her ability to stand her ground makes her a refreshing and likable heroine.
Familiar faces from The Deal, including Garrett and Hannah, return, while new side characters add to the college setting’s energy and humor.
Kennedy’s writing is engaging, with a mix of humor, emotional moments, and steamy romance. The alternating perspectives between Logan and Grace allow for a well-rounded view of their relationship and personal struggles. The book delivers plenty of humor and flirty banter while also tackling deeper themes of family responsibility, self-worth, and making tough life choices. The chemistry between Logan and Grace is strong, and their romance is a blend of fun, tension, and genuine emotional connection.
Strengths
✔ Strong Character Growth – Both Logan and Grace undergo realistic, satisfying development.
✔ Second-Chance Romance Done Right – Logan’s journey to making up for his mistake feels earned, and Grace doesn’t immediately forgive him, which adds authenticity.
✔ Engaging Writing & Humor – The witty dialogue and college setting keep the story entertaining.
✔ Balanced Romance & Personal Struggles – The love story is front and center, but each character also has their own challenges to overcome.
The Mistake is a fun, emotional, and steamy addition to the Off-Campus series. While it follows a familiar formula, the engaging characters and well-paced romance make it a standout. Fans of The Deal and other college sports romances like The Chase by Elle Kennedy or The Score (book 3 in the series) will love this one.
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Reviewed in the United States on February 16, 2025
★★★★★ 5
Love this cleanser
Size: 13.52 Fl Oz (Pack of 1), Style: 13.52 Fl Oz (Bottle)
I really love their products. This one in particular stands out to me bc I get oily around my chest and get little superficial pimples on my arms/neck and back and WOW!! This stuff really gets them gone! I use this all over my body the oily parts and I just love it! No smell at all. Very gentle. Not greasy! Great deal for the size and it’s been a game changer. I’ve realized I’m sensitive to even sensitive skin cleansers and this one doesn’t irritate at all!
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Reviewed in the United States on May 21, 2026