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Description
Mouse ANXA7 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 Annexin A7 (ANXA7) 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 Annexin A7 (ANXA7) content 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 A7 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Annexin A7 (ANX/A7) is a protein encoded by the ANXA7 gene. It is a member of the annexin family of calcium-dependent phospholipid-binding proteins. The gene contains 14 exons spanning approximately 34 kb of DNA. An alternatively spliced cassette exon results in two mRNA transcripts of 2.0 and 2.4 kb, predicted to produce two protein isoforms with different N-terminal domains. Alternative splicing events are tissue-specific, with mRNAs containing the cassette exon prevalent in the brain, heart, and skeletal muscle. Transcripts also differ in their 3' UTR by the use of two alternative poly(A) signals. The choice of poly(A) signal is independent of the mRNA splicing pattern. It has been shown to interact with ALG2 and SRI. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.2 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 4
I'm a Total Goner
Format: Kindle
🦇 The Fiancée Farce Book Review 🦇
Rating: ⭐⭐⭐⭐
❝ She was a total goner. Whatever magic Tansy was made of, Gemma wanted to drown in it, revel in the honeyed heat burning her up from the inside out. It was better than the finest bourbon she’d ever had the pleasure of sipping. ❞
🦇 After losing her parents and a traumatic romantic experience as a teen, Tansy Adams focuses all her time and energy on her late father's bookstore. To dodge questions about her love life, Tansy invents Gemma, a fake girlfriend inspired by the stunning cover model on a best-selling romance book. She never expected that real-life Gemma would step into her life, play along, and announce they were engaged, imploding that white lie into a full-on farce. Gemma van Dalen, the outcast of her wealthy family, needs a spouse to inherit Van Dalen Publishing; her grandmother's legacy. In exchange for Tansy's hand in marriage, Gemma offers to save Tansy's beloved bookstore before it's sold off. Can their marriage of convenience work, or will Gemma's scheming family get between them?
❝ "As for feeling like you’ve failed, well, failure is an inescapable part of life. But failing doesn’t make you a failure. And I’m sorry your father made you feel like the two were synonymous. You are more than your achievements, Gemma. You are brilliant, and ambitious, and you are good, do you hear me? And what you do or do not achieve in this life has no bearing on your value.” ❞
💜 Oh. My. Goddess. That's right: goddess, because that's exactly what these two cunning, sassy women are. Alexandria Bellefleur has taken the generally predictable "marriage of convenience" trope and turned it into an emotional, powerful story about finding the love you don't realize you deserve. Both Tansy and Gemma are full of so much pain, their family trees full of broken branches (and in Gemma's case, poisonous barbs). For them to find love and family in one another so unexpectedly (for them, at least, because, come on, that's what we're here for), so flawlessly...it's not only swoon-worthy and sweet, but a relief. It's the insane, instant, undeniable chemistry between Tansy and Gemma that empowers their every interaction from the start, but unlike other marriage-of-convenience stories, smut doesn't drive their relationship. What starts off as a business relationship blossoms from a friendship to a true partnership. Gemma is sweet and giving as she navigates her first real relationship, while Tansy comes out of her shell to defend Gemma when no one else has. These goddesses support one another, even when the relationship is only a farce, until it all becomes real. The character development between them both is a flawless example of how empowering love--and having someone by your side--can really be.
🦇 As much as I loved every interaction between Gemma and Tansy, it's difficult to love everything that happens outside of their sweet sapphic bubble. The toxic men--namely Tucker, Gemma's cousin, who manipulated teenage Tansy and shared underage nudes of her--seem unrealistically cruel. Tansy's step-mother goes from a social ladder-climbing step-Bridzilla to suddenly sympathetic. None of the secondary characters have real layers, making them no more than pawns to the story's plot progression. Because of that, none of the conflicts or twists are surprising. The boardroom scene/resolution seemed beyond unrealistic, regardless of how sweet the gesture was. You may need to expand your suspension of disbelief and focus on the sapphic sweetness for this one.
🦇 Recommended for fans of Stars Collide, Cleat Cute, and Love at First Set. This heartwarming sapphic romance is full of feels; a stunning addition to any shelf.
✨ The Vibes ✨
🩷 Fake Dating
🩷 Marriage of Convenience
🩷 Bi MCs (Bi4Bi)
🩷 Sapphic Romance
🩷 Mental Health Rep
🩷 Opposites Attract
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Reviewed in the United States on July 10, 2024
★★★★★ 5
The BANTER 😍
Format: Kindle
Thoroughly enjoyed this book. The perfect witty, banter-laden escape, even if predictable for me, which isn’t even that part of the comfort and fun?
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 9, 2025
★★★★★ 3
okay
Format: Kindle
I’m honestly not sure how i finished this book. i considered dnf-ing multiple times due to boredom, and then did that cycle like 5 times??? So here are your stars, i guess.
The plot is bonkers, and somehow was still sooooooo boring to me. Every single twist was painfully obvious, and the chemistry between Tansy and Gemma was saccharine in a very cloying way. The best way i can think to describe this book is having a trope wheel in front of you, spin it 5 times, and write that down.
I didn’t hate my reading experience, but i also didn’t *enjoy* it.
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Reviewed in the United States on June 9, 2024
★★★★★ 5
Another Fantastic Story From Alexandria Bellefleur
Format: Kindle
Love this author so much that I THOUGHT I had pre-ordered this book. Now I'm late to the party, but realizing that this book had been released made my day yesterday. Less than 24 hours later, I've finished the book and I miss it already.
In my opinion, Bellefleur is damn good at writing romances that feel like they could be real. That's not to say they aren't swoonworthy - they definitely are - but she finds the romance in the everyday instead of relying on overly flowery declarations and grand gestures. Okay, fine, there are some grand gestures, but it's the combination with the little things that make her love stories so great.
A couple quick additional things I will say about Bellefleur's writing at this story:
- She stays true to her characters. They change and grow, but they do so in a way that fits them or aligns with the influence of their partner. I also appreciate the fact that her characters don't talk and act the same way from book to book. Each character feels like someone new to get to know.
- The spicy scenes are - *chef's kiss* - practically perfect. A good amount of scenes with a good amount of words dedicated to each without any cringeworthy metaphors.
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Reviewed in the United States on May 2, 2023
★★★★★ 4
Cute!
Format: Kindle
This was really cute but once I got to around 60-70% I felt like the story dragged and it took me much longer to finish this than was necessary. Also idk if it was me, but for some reason the way the characters talked at times reminded me of a Historical Romance novel and it had me ???
Still a good time and I'm glad I read it :)
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Reviewed in the United States on October 9, 2023
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