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Description
Mouse EGFL7 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 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. 3. 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. 4. 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 an EGF-like domain-containing protein 7 (EGFL7) 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 EGF-like domain-containing protein 7 (EGFL7) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse EGF-like domain-containing protein 7 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Epidermal growth factor-like domain-containing protein 7 (EGFL7), also known as EGF-like domain-containing protein 7, is a protein encoded by the EGFL7 gene. Its intron 7 contains the miR-126 microRNA gene. In vitro, it inhibits PDGF-BB-stimulated aortic smooth muscle cell migration but has no effect on cell proliferation, suggesting a role in vascular maturation. In vivo, knockdown of EGFL7 expression in hepatocellular carcinoma cells reduces the number of intrahepatic and pulmonary metastases. Within tumors, EGFL7 increases microvessel density, hypoxia, necrosis, and vascular permeability. It is a natural negative regulator of vascular elastogenesis. It interacts with and inhibits the catalytic activity of LOX, preventing the cross-linking of elastin molecules into mature, insoluble elastin. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenates and other biological fluids |
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4.3 ★★★★★
Based on 2456 reviews
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Product Reviews
★★★★★ 5
So good!
Format: Kindle
Wings of Stars was a fantastic first book to a new series. It grabbed me in its clutches within the first chapter and kept ahold of me.
This new world of M Sinclair and RL Caulder’s is all about angels and mythical creatures. It’s full of drama from all points. From Kieran’s awful home life to keeping the stars alive and everything in between.
Kieran is our FMC and despite everything she keeps her spirit and defiance.
Keeping to their norm, there is a group of guys for our FMC but they definitely aren’t established yet, and not fully cemented yet into being a group rather than her choosing between them.
Ronan is so sweet and gives off Daddy vibes
Gabe is also sweet and wants the best of everything for Kieran and will defend her against anyone
Bastian is slightly unhinged but I loved him immediately
Steele is a jerk… just as you find redeeming qualities, he screws up again… but yet I still want him to redeem himself.
There’s another one… but if I gave his name I would be spoiling a plot twist so you will just have to find out yourself… but you’ll love him as much as I do.
This is a slow burn but the little bit of spice we do get is hot, especially with the dirty talk.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 19, 2024
★★★★★ 5
Angels and the Fallen — Breathtaking Start
Format: Kindle
This book was EVERYTHING I expected it to be and more. Leave to R.L. Caulder and M. Sinclair to give us yet another amazing book!
Kieran and her guys hooked me from the very beginning and did not let go. Found family is one of my favorite tropes and it was done so well in this book. Couple that along with an FMC trying to find her place in the world and I was practically drooling over this book. The world building, the conflict, the wyverin sidekick — it all was done so well that it felt fresh and real.
Each love interest felt real and unique in their own ways. I felt connected to each one and like they were truly different people. I love that so much in a book and these two authors never miss with their love interests.
Multi POV, Reverse harem, who did this to you, and a magical world highlighting the angels and the fallen. This book has everything and it does it so well.
Big thank you to R.L. Caulder and M. Sinclair for the arc copy! I cannot wait for book 2!
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Reviewed in the United States on February 20, 2024
★★★★★ 4
A new take 🤔
Format: Kindle
I’m not going to go into what this book is about — by now you e already read the synopsis and have an idea — but I will give a couple of insights.
Definitely a different take on angels and their fall; I like it. This is an interesting start to a new series - YES, this does end on a cliffhanger so be prepared for that. I’m eager for the next book (and hopefully the last in this series — sometimes the author(s) will switch it up depending on how the story flows for them) in this series.
I’m hoping that a certain male character redeems himself because he’s a dick that will make the meaning of mixed signals jealous lol.
You ever watch that old movie — I think it’s call Mommy Dearest? — yea, let’s switch that up to daddy. That man would make a perfect husband to mommy dearest.
I recommend this book because while these authors are well known for their cliffhangers, they are also well known for putting out good stories.
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Reviewed in the United States on February 19, 2024
★★★★★ 3
Liked it
Format: Kindle
I liked the worlds and the characters. The prophesy is confusing in that I would think, since all worlds ore on a path to destruction, the angels would want their world to be saved. I am sure there is more evil that will be revealed as the series proceeds. I dislike a prolonged “poor little me” trope and the main character has moved past that, thankfully, and began to show a fierce attitude as the novel progressed. The last portion of the novel had some great action. It’s worth reading, just be patient with a slow start.
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Reviewed in the United States on February 14, 2024
★★★★★ 5
AMAZING debut novel!!!
Format: Kindle
Plot ⭐️⭐️⭐️⭐️
Spice 🌶️🌶️.5
Romance 💘💘💘
Vibes ⭐️⭐️⭐️⭐️⭐️
Dual 1st person POV - Ara (26) & Rogue (39 - but looks mid-20s: they can live hundreds of years so this isn't that large of a gap as it could've been which I heavily appreciate lol)
Tropes: enemies to lovers, fae/human wars (deep hatred for each other), shifters (dragons- MMC can only partial shift with wings), one horse, one bed, touch her and d!e, found family, abduction turned to freedom
The Last Storm is the debut novel from JD Linton and let me tell you, you guys NEED to read this. The plot was engaging and the editing was was amazing (especially for a debut novel). Our FMC, Ara, is stuck in her gilded cage longing for a life outside of her small town. She uses her books to escape and live vicariously through the pages (honestly, relatable). After her father announces her betrothal to her childhood friend (to whom she has no romantic feelings for), Ara tumbles unknowingly into a desperate plot trying to stop the humans from slaughtering the Fae.
As one can expect from an enemies to lovers / kidnapper/captive romance, Ara fights her attraction and lust towards our MMC, Rogue (the King of the Fae), for as long as she can. Upon seeing Ara for the first time, Rogue is instantly aware that she is his fated mate (not a spoiler). Since she is the General's only daughter, he plans to abduct her and use her as leverage to stop the brutality. During Ara's time in Rogue's captivity, their banter and chemistry continue to rise until they finally boil over and come together (quite literally, and many times I may add 😉).
Here's what I LOVED:
- Rogue continuously seeks advice from his elders and deeply respects their opinions and life experience and tries to implement their recommendations
- Rogue makes many mistakes in the beginning but we see him actively work on not repeating them as the book progresses. The level of self-awareness and his ability to change his behavior was impressive
- The magic system is intricate and we have only scraped the surface. As the series continues and Ara progresses in her powers, I'm sure we'll get to see more of this. I absolutely LOVE the messaging system that is used in this book.
- Ara's struggles are so human and so raw. She is experiencing so much guilt and pain and hurt and getting to see her work through each of these emotions is inspiring. Especially as her and Rogue get closer and she learns she can lean on him as well, that she is not alone.
- While this is the start of a series, there is NO cliffhanger! There's a bit of a teaser of something major that is going to happen at the start of the next book, but it's not a cliffhanger in the sense that we aren't sure if someone is going to live or d!e or if they'll be separated. For that, I am very thankful! This book was so much fun that I will definitely be returning to book 2, even if it takes several months (or longer since this is an debut author) to publish!
- Lastly, the cover is GORGEOUS! And I love the title!
I'll copy a few of my favorite quotes below so you can have a little taste of the author's writing and the world she's cultivated. 😊
Top Highlights from The Last Storm
On days like this, when my heart was heavy and my mind clouded, I resorted to books— to escape, to forget, to find freedom where I had none.
If I were to marry him, my face would always be turned to the window, searching for more, and if not that, I would be a shell of the person I am now.
I stepped back to admire her, thr0bbing at the sight. She was the most beautiful woman I had ever seen. To ever exist. Nothing, no one, had ever deserved to be worshiped more. All men should be made to kneel before her. But she would have to settle for me.
The taste of her met my t0ngue as my scent merged with hers, forever branding her. Mine. I l!cked the wound. Hers. Completely and utterly hers.
I didn’t claim her in ownership. I claimed her as my one. Devoted myself to one. With that mark, my body and soul were bound to her. I would never be with anyone else, emotionally or physically. It would be her or no one, until my last breath.
“Scream my name. Let everyone know who I belong to.”
I had never really cared about the weather before, but now, clear skies meant everything to me, and I was grateful to see another calm morning.
“There will never be another woman for me.” He paused. “Ever.” I stilled at his words. “What… Why?” “This”— his thumb slid down across the mark—“ is a symbol of… surrender. I know you believe that it was my claim upon you, but it wasn’t. It never was. I bound my body and soul to you, little storm.”
“I also know that it is more than this tiny, insignificant mark on your skin that binds me to you. It’s you. All of you. Your strength and resilience. Your determination to endure no matter what fate throws at you. Your love for love and stories and hope. You are entirely the opposite of everything that I am and I would gladly wear your shackles if it meant I could have you.”
My mate. Mine. And then everything shifted and I understood. I understood everything. The surrender. The deep, soul-craving longing. Bound. I was bound to him. Body and soul. Entirely his. “I would’ve waited forever,” he whispered back, understanding.
Seriously, everyone.. add this to your TBR!!
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Reviewed in the United States on November 14, 2022