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Description
Mouse ANXA1 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. 5. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 2000pg/mL). Then dilute to the following concentrations: 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 2000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 1000pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 enzyme conjugate are sequentially added to microwells pre-coated with an Annexin A1 (ANXA1) 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 level of Annexin A1 (ANXA1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Annexin A1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Annexin A1 (AnxA1), also known as lipocortin I, is a protein encoded by the ANXA1 gene. It belongs to the annexin family of Ca2+-dependent phospholipid-binding proteins with a molecular weight of approximately 35,000 to 40,000 daltons and is preferentially localized to the cellular surface of the plasma membrane. With an apparent molecular mass of 40 kDa, it inhibits phospholipase A2. It has important opposing properties in innate and adaptive immune responses: it suppresses innate immune cells and promotes T cell activation. T cell activation leads to the release of Annexin A1 and the expression of its receptor. This pathway appears to fine-tune the intensity of TCR signaling. Under pathological conditions, elevated Annexin A1 expression may increase the intensity of TCR signaling through the mitogen-activated protein kinase pathway, leading to a hyperactivated state of T cells. | |||||||||||||||||||||||||||||||||
| 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 | 31.2-2000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.8 ★★★★★
Based on 14 reviews
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Product Reviews
★★★★★ 5
What did i spend so much time reading??
Format: Kindle
Spoilers ahead This book makes me angry. A misogynistic fable that holds women as mothers as inherently either deeply flawed or as if some fairy tale perfect mother exists. I found the early parts of the novel quite funny. Thinking this poor woman why do people expect such crazy standards? Also social media influencers somehow we are to take for reality? It is entertaining and that's why we want to spend our time looking at it. It is a fantasy world. Yet that is somehow turned from something comical and poignant dissolving into madness where there can be a 16 year old that can take away your children for what I'm not sure. Filming them? Having nannies? And then again the next bunch for living off the grid? I get that this isn't going for realism. It ultimately became for me this poorly written satire maybe? Obviously what started as comically intriguing for me descended into one totally crazy crazy turn to the darkest of places. I recognize this was supposed to be some sort of cracked fairytale where nearly everyone is evil and/or mental but 30 years for child abuse? This is like a nightmare not a thriller. The book demonstrates a hatred for mothers. Just awful! It made me crave for a totally unrealistic thriller. I came to be entertained and left pissed off.
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Reviewed in the United States on May 13, 2026
★★★★★ 5
Excellent satire of tradition cult
Format: Kindle
I read this book in two days. I was eager to read it once I read the description in a Facebook post. I am a theologically conservative Christian frustrated with the tradition culture that has co-opted the faith. I see this in my social and social media circles quite a bit, and I was intrigued by a novel that took this movement to task.
Furthermore, I truly enjoy well done satire and farce. I have had a hard time finding these works written well in the past, so I was bracing myself to be disappointed. I am thankful to say, I wasn’t. I didn’t expect the novel to go the way it did (and no, I won’t post spoilers), but I literally laughed out loud a several points.
Let me compliment the author for one particular point: She doesn’t just blast the “tradwife” movement. Plenty of cultural movements and groups of people have their sins exploited. Rather than shooting a 12-gauge at religious conservatives, she takes a Tommy gun and hits a wide range of systems.
I do want to warn squeamish readers about another issue: The novel can be quite graphic at times, explicitly detailing the intimacy struggles with the main couple. It borders on the pornographic side, but not for titillation. It’s meant to show the dysfunction in the supposedly happy couple.
There were quite a few humorous moments, such as the protagonist enjoying the sounds of the chickens, only to grow wistful about their impending deaths so she can taste chicken broth. Or the protagonist’s evangelical mother reacting to her son-in-law’s purchase of a yoga mat.
I was a little disappointed in the resolution of the central issue, i.e., the time travel. But overall, the novel was an enjoyable read and worth your time.
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Reviewed in the United States on May 8, 2026
★★★★★ 4
What did I just read
Format: Kindle
I’m not sure how to articulate my feelings about this book. It took me a bit to get into it, the story was interesting from the beginning but felt very slow paced to me. That changed about half way in. I don’t know what I was expecting, but that was not it! I did end up enjoying this. It was well written, funny, and shocking.
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Reviewed in the United States on May 30, 2026
★★★★★ 5
Absolutely the best in the series so far!
Format: Kindle
I've been wanting to get Dean's book since we met him in The Deal, and let me tell you, this book did NOT disappoint!
Dean is the playboy of the hockey team, he doesn't do relationships, and he just takes life as it goes. Allie is Hannah's best friend and roommate, and is a relationship type of girl. When she realizes her and her boyfriend are moving in separate directions and she can't do it anymore she breaks up with him. When she needs a place to hide out, Garrett offers up his house while they are away. She never expected that weekend to change everything for her.
Dean and Allie definitely have amazing chemistry, but one night together doesn't change things for them, or does it? Dean can't get Allie out of his head, and Allie knows she can't do a casual hookup, but they can't seem to keep away from one another. I loved how Dean finally wore Allie down, and how Allie finally got to have someone with her that understood her and was just as adventurous as she was. I don't agree with how she didn't want anyone to know about them, but I think the way everyone found out was perfect for them.
Dean and Allie's "relationship" is full of emotions, and you can slowly see this turning into something way more than what they thought it would be. Dean is amazing with Allie, he's supportive of her, he cares about her, and he's insanely jealous of anyone who dares look at her. Allie is loving how Dean makes her feel, and is also a huge supporter of his. For once in his life Dean opens up about himself and you can tell how nervous he was telling her about his ex, and everything involving the fall out of that situation. Their escapades are hilarious and sexy all at once. I loved watching them during Thanksgiving with Beau and his sister, where you can see that things definitely changed for them. I also adored their watching of the french soap opera, and all the little things they did together.
Not only does this book have a true romance that you can't help but fall in love with these amazing characters, but there are some absolute laugh out loud moments. When Logan walks in on Dean and "Winston" I spit my drink out I was laughing so hard. Also the hilarious Twilight references on imprinting! There is also a very serious aspect to this book that had me in tears. My emotions were definitely all over the place while reading this book, and I must say I think I love Dean more than Garrett, and he's been sitting at the top of my BBF list for a long time!
It's been a long time since I've read a book that I absolutely loved both of the main characters. I felt so connected to them, and their group of friends, that I thought I was right there with them experiencing their love, their laughs, and their heartbreak. This book definitely took me on one hell of a ride. I can't remember laughing so hard one moment, swooning and falling in love, and then balling my eyes out, only to have my heart put back together at the end. This is a beautifully written story and I could NOT put it down. I am definitely looking forward to Tuck's book, especially with the bomb-shell he dropped at the end of Dean's story!
Even though this book can be read as a stand-alone, I highly suggest reading each of the previous two books in this series, so you can get a good feel of how much Dean has changed since we met him, as well as how the group dynamic works. Each one of the books are 5 star reads, and trust me, you will definitely fall in love with Garrett, Logan and Dean!
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Reviewed in the United States on January 12, 2016
★★★★★ 4
The Good, The Bad and Everything In Between
Format: Kindle
Another excellent entry: The Off-Campus series is among the very, very best in new adult romance and there is little that isn’t done well in this entry.
Dean DiLaurentis is charged with watching out for Allie, Hannah’s roomie, one weekend while everyone is gone for various reasons - with, of course, the obligatory threats to his penis if he were to touch Allie. She’s just gone through a painful breakup with her boyfriend from book one, since they’ve had a fairly tempestuous relationship, she needs some time away from her apartment, where he’s been trying to get her to change her mind.
Of course, Allie and Dean are young, hot, single and there’s tequila. When you do the math, it’s fairly obvious that yes, Slot A goes into Tab B, and yes, romance happens. I love their banter, the humor throughout really establishes these characters and their friendships and makes the ride all the more enjoyable. Speaking of riding...
Dean’s what I expected: Dean’s the manho who beats all the previous manhoes in this series. He makes no apologies and I loved that he didn’t. He’s intrigued with Allie but he isn’t in love from the moment he lays eyes on her. When he falls, though it’s hard and it’s clearly life-altering. His moments with his friend Beau when they discuss his issues with little Dean (I can’t stop giggling at that silliness) were priceless.
He has a good journey towards adulthood, though I was saddened by the events that led to this character growth. It was the perfect (sad yet realistic) touch of bitter to make the sweet all the richer.
Allie was a surprise: I’ll admit, this is mostly because I didn’t remember her. Allie isn’t a big part of the first book and what I did remember of her was a bit flaky. I loved getting to know her and seeing what made her tick and as with Ms. Kennedy’s previous heroines, there is more than meets the eye. She’s concerned about being slut-shamed by the prospect of no-strings sex, considering she’s only ever been in committed, long term relationships, so I love that she has to learn to accept that there’s nothing wrong with two consenting adults doing what they want to do, damn public opinion. I found it an interesting twist that the only person truly slut-shamed is Dean, by other men and women.
Beats played out as I expected: If I have any issues with this is, while it is entertaining, charming, and funny, it also felt the same as the earlier books, which were also entertaining, charming and funny. When the quality is so high, that’s not a horrible thing, but I can’t help but struggle to keep each set of heroes and heroines separate in my mind. That said...
I’m really intrigued by where this series is going next: The setup for the next book is markedly different and it has me really eager to see how Ms. Kennedy handles it. The heroine for the next book is NOT in the mold of the earlier women and the hero is a bit of a wild card, since he has been more on the periphery so far. I’m looking forward to seeing what they have to contribute to this universe.
The Bottom Line
This is an excellent new adult romance with likeable characters, funny banter, hot sex scenes and room for the ‘verse to grow. If you haven’t read Ms. Kennedy’s series, I highly recommend it.
**ARC provided by author for review**
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Reviewed in the United States on January 14, 2016