SKU: 57302545899

Human Anxa1 ELISA Kit

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Description

Human 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. 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.
5. 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.
6. 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.
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 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 Human
Synonym Human Annexin A1  ELISA Kit
Detection Type Double antibody sandwich method
Composition
Name 9 6 T  match   set remark
Pre-coating 96 Well plate 8 Hole ×12 Strip without
Standard 2 branch
Dilute as per instructions
Universal diluent
2×20mL
without
Concentrated biotinylated antibody ( 100× )  
120uL
Dilute as per instructions
Concentrated enzyme conjugate ( 100× )
120uL
Dilute as per instructions
20× Washing liquid
2×10mL
Dilute as per instructions
Bottom thing ( TMB )
10mL
without
Stop liquid
6mL
without
Sealing film
4 Zhang
without
Instructions
1 Share
without
Background Annexin-A1 (Anxa1), also known as lipocortin I, is a protein encoded by the human ANXA1 gene. Annexin-A1 belongs to the annexin family of Ca2+-dependent phospholipid-binding proteins with a molecular weight of approximately 35,000 to 40,000 daltons. It is preferentially localized to the cellular surface of the plasma membrane. Its apparent molecular mass is 40 kDa and it inhibits phospholipase A2. Anxa1 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 Anxa1 and the expression of its receptor. Under pathological conditions, elevated Anxa1 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.
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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SKU: 57302545899

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4.5 ★★★★★
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Product Reviews
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Britt
Louisville, US
★★★★★ 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
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Kindle Customer
Chelsea, US
★★★★★ 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
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Verified Purchase
Jennie_reader
Battle Creek, US
★★★★★ 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
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shellbelle
Draper, US
★★★★★ 5
I loved everything about The Deal by Elle Kennedy!
Format: Kindle
I have been a huge fan of Elle Kennedy’s writing for years. I loved her Out of Uniform series and Killer Instincts series. Her After Hours series is sizzling hot. She has a way of bringing her characters to life and making them demand your absolute attention. Her books have a combination of witty dialogue, dynamic characters, a well developed plot, and pacing that keeps the reader enthralled. Most of her books have either been erotic romance or romantic suspense. I was wondering how she would pull off a New Adult romance. New Adult romance is a generalized genre that can contain sweet & sexy, dark & intense, angsty & anxious, or exciting & erotic. The Deal has all of these elements and more. It is one of the most enjoyable and entertaining New Adult romances that I have read. I connected with the main characters as well as the secondary characters. They were all larger than life in their own way. The one thing that stood out besides the main characters romance was Elle’s portrayal of college life. She portrayed a very realistic look at life on the college campus and how it relates to their courses as well as social life. Garrett Graham is the bad boy of the college hockey team. He makes panties melt and owns his man whore reputation with relish. He has an overinflated ego that matches his cocky attitude. He is a star player and lives for playing hockey. He is diligent with his grades, practices, and performance. When he fails his philosophy mid term, it could not only ruin his GPA, it will affect his position on the team. He could lose the one thing he lives for. He is going to have to retake his philosophy mid term and ace it in order to remain on the team. He needs help. Who better to help him than one of the few students that managed not only to pass the mid term but ace it. He never really noticed Hannah Wells before until she accidentally dropped her exam and he saw her A. Now he is on a mission to get Hannah to tutor him. He will try to bribe her, entice her, and woo her. When he discovers the golden carrot that will make her give in, he will dangle it in every way he can. His life is counting on that golden carrot. Hannah Wells is a music major. She is a pretty confident and outgoing girl with her friends and peers. She is smart, personable, and resilient. She is dedicated to her craft. The one area she lacks in is her confidence with the opposite sex. She has finally found a guy that she is interested in and she has no clue how to deal with her attraction. Not only is he out of her league, he is also the star quarterback on the football team. Not the type of guy she would normally take an in interest in. She has had some very bad experiences with the opposite sex. Although she has mostly overcome her issues, she is still a bit leery and scared. If she can get past her fears, she may be able to approach him. When the ego maniac sex god, Garrett Grant, approaches her to tutor him in philosophy, she thinks he is out of his ever loving mind. There is no way she is going to tutor the self absorbed jock. That is until he finds out her ultimate weakness and offers her a guaranteed solution. His persistence and golden carrot are enough to make her cave. It’s time to make a deal with the Devil. Hannah and Garrett are in for some big surprises. They are both quite different than they appear on the surface. Hannah may have the upper hand when the tutoring begins but Garrett will quickly take control of the situation. Both will be in for a great learning experience. Both will come out winners in the end. Their Deal will seal both their fates. I loved the banter between Garrett and Hannah. The snarky sarcasm, witty comebacks, and dirty talking are beyond clever and entertaining. The heated bedroom scenes make your temperature burn out of control. The emotional and intimate moments will have you swooning. What I loved best is how they formed a deep respect and friendship with one another. They were able to honestly confide in one another and know that their trust was not misplaced. I loved everything about this book! Read The Deal by Elle Kennedy. I am looking forward to the next book in the Off-Campus series. The Mistake will release in June of 2015. It is a complete mystery of who this book will feature or what their story is about.
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Reviewed in the United States on March 23, 2015
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Tori
Boise, US
★★★★★ 4
A Beloved College Romance That Earns Its Hype.
Format: Kindle, Format: Kindle
The Deal by Elle Kennedy is a college romance that fully delivers on its reputation. With engaging characters, steady emotional growth, and a relationship that develops naturally, it’s easy to see why this book remains so popular. The story follows Hannah Wells, a focused and reserved music student, and Garrett Graham, the confident captain of the college hockey team whose academic struggles threaten his future. When their paths cross through an unexpected agreement, a friendship begins that gradually deepens into something more meaningful. One of the book’s greatest strengths is its pacing. The relationship doesn’t rush or rely on instant attraction alone. Instead, it builds through shared experiences, honest conversations, and growing trust. This approach makes the connection between the characters feel believable and emotionally grounded. Both Hannah and Garrett face personal challenges that shape their decisions and growth throughout the story. These elements add depth without overwhelming the romance, keeping the focus on character development and mutual support. The dialogue is lively, the humour is well-placed, and the emotional moments are balanced with lighter scenes that keep the story moving. While some plot elements may feel familiar to readers of the genre, the overall execution is strong. The conflicts fit naturally within the story, and the resolution is satisfying without feeling overextended. The Deal is an enjoyable, well-written romance that blends heart, humour, and character-driven storytelling. A solid choice for readers who enjoy college settings, slow-burn relationships, and emotionally engaging narratives.
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Reviewed in the United States on January 18, 2026

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