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Description
Human Kindlin-2 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 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 a kindlin-2 capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of kindlin-2 in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Kindlin-2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Fermitin family homolog 2 (FERMT2), also known as pleckstrin homology domain-containing family C member 1 (PLEKHC1) or kindlin-2, is a protein encoded by the FERMT2 gene. Kindlin-2 was the first kindlin protein discovered in 1994. It was discovered during a screening for epidermal growth factor (EGF)-induced mRNAs and was originally named mitogen-induced gene 2 (Mig-2) protein. | |||||||||||||||||||||||||||||||||
| 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 homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.3 ★★★★★
Based on 2483 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.
WAS THIS REVIEW HELPFUL?YesReportShare
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.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 30, 2026
★★★★★ 5
Meaningful, Enjoyable and Uplifting
Format: Paperback
I don’t read a lot of novels, but after hearing recommendation after recommendation, I finally picked up Theo of Golden—and it absolutely lived up to the hype. Alan Levi tells a story that is inspiring, wholesome, thoughtful, and genuinely fun to read. The characters felt real, the writing was engaging, and the story carried deeper themes about life, purpose, and relationships without ever feeling forced. I also know Alan through Young Life, which made reading the book even more special for me personally. If you’re looking for a meaningful novel that is both enjoyable and uplifting, I highly recommend this book.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 23, 2026
★★★★★ 5
a swim through literary magic
Format: Kindle
Reading this book reminds me of jumping into a pool of water and the way the water tickles your skin as you descend into it, then surrounds you and totally encompasses you in its liquid wonder.
There is so much here: beautiful metaphors that conjure up both visual images and the emotions that accompany it. Wonderful character development: presented with gentleness, respect and grace. An intricate plot that carries the reader along lightly and buoyantly, not calling attention to itself, but carrying through to the end. The book itself is a portrait of Theo, in keeping with the portrait style described within.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 21, 2026
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