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Description
Mouse SPTAN1 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 100 ng/mL). Then dilute to the following concentrations: 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.125 ng/mL, 1.5625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 100ng/mL standard working solution into the first EP tube and mix thoroughly to make a 50ng/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 utilizes 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 Alpha-Fodrin (SPTAN1) 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 Alpha-Fodrin (SPTAN1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Alpha-Fodrin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Alpha II-spectrin (α-fodrin), also known as spectrin alpha chain, is a protein encoded by the SPTAN1 gene. Alpha II-spectrin is expressed in multiple tissues and is highly expressed in the myocardial Z-disc, costameres, and sarcolemma membranes. Alternative splicing of αII-pectrin has been documented, resulting in multiple transcript variants. Specifically, four αII-pectrin splice variants have been identified in cardiomyocytes. In contrast to αII-pectrin, which is primarily present in erythrocytes, αII-pectrin is expressed in most tissues. In cardiac tissue, αII-pectrin is found in the Z-disc, costameres, and myocardium of myocytes, as well as along the cytoskeletal network of cardiac fibroblasts. αII-pectrin most commonly exists as a heterodimer with αII and βII-spectrin subunits. Dimers often self-associate and heterotetramerize. | |||||||||||||||||||||||||||||||||
| 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 | 1.56-100 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.1 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 5
Quality and security-oriented wallet
Color: Adventure Brown - Forest Brown
This is a neatly constructed wallet. The material of the leather feels satisfying to carry unlike previous wallets. This wallet consists of many card slots, and allows ease of access between them. The wallet can fit checks, cash and is just a nice layout and style to be able to look through it. Because some of our cards support RFID and NFC technology, bad actors can try to steal our credentials. This wallet has a material to mitigate these occurrences, but like anything in security whether it be physical security controls or the wide scope of cybersecurity, do not rely on it and try to be up-to-date with the trend. I am sure we would all want to be one step above in relation to keeping our credentials secure. Still, it is an excellently built wallet and I highly recommend it.
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Reviewed in the United States on May 7, 2026
★★★★★ 5
Easy to use
Color: Adventure Brown - Forest Brown
Cards load easier and easy to remove them to use. Holds money and cards for easy use. Nice and supple and wears excellently. Good quality leather and card slot are bigger then other wallets I have tried.
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Beautiful Wallet
Color: With NO Logo - Smooth Nappa Black
Well constructed and beautiful wallet. The black Nappa leather I ordered is soft and smooth to the touch and you can tell by the look, feel, and smell that it is high quality. I love that you can order this with both a logo and no logo for a clean look.
The design on this is well thought out.
I'd definitely recommend this wallet to anyone. It is an amazing value and far superior to any other $30 wallet I've ever seen.
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Reviewed in the United States on March 19, 2026
★★★★★ 4
Almost perfect!
Color: With Coin Pocket - Forest Brown
I purchased this wallet primarily for its generous number of conveniently located slots and its coin pocket. If you can't fit everything you want to EDC in this wallet, you need to think about getting a sling bag! Having returned the same model in Grizzily Brown due to a stitching defect and ordered this one in Forest Brown, I can see it is a richer, lighter shade and the finish seems smoother and more refined. Some of my cards were tight in previous wallet slots, but everything fits great in this one. Why the one star off? First, this wallet is about a quarter inch too large in all dimensions, especially thickness. While it doesn't protrude out the top of my back pocket, it is a challenge to close the button on some pants and it is also snug side-to-side, so removal involves some wiggling until pockets stretch and the wallet breaks in. The leather used in its construction is smooth but stiff at first, given that it is about twice as thick as most American or European wallet leather I've owned: think bullhide. The smell is also a little off, definitely natural cowhide but processed with whatever is used to tan leather in India rather than in the United States. Finally, this looks like something that belongs in a pair of jeans or casual pants, NOT pulled out of a suit at a Michelin star restaurant. The stitching, evenness of dye and lines are far from perfect, but what it lacks in looks it more than makes up for in every day function, character, and anticipated durability.
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Reviewed in the United States on January 13, 2026
★★★★★ 5
The Best Wallet
Color: Boulder Black - Napa Smooth Black
This is the second wallet I have purchased from Bull Guard. The first one is identical and I have used it for approximately one year. The first wallet has held up perfectly. I just decided to buy a second one because I am so happy with the first one and I decided to break up my cards into frequently used in the second, new wallet and less frequently used in the first and older wallet. The capacity of the wallet is excellent. It easily holds all my credit cards, drivers license, bills, insurance ID cards, and my global entry card. The material, workmanship, durability, and quality is also excellent. The wallet is all leather and breaks in perfectly. The size is also perfect fitting perfectly in my rear pocket. For the price, this is the best man's wallet I have ever had and I highly recommend it.
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Reviewed in the United States on November 22, 2025
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