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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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4.2 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
Perfect Small-ish Business Bag
Color: Apricot, Size: 14-Inch
I have been on the search for a small business bag, that can go on. top of my luggage as well as look nice in the office and at meetings. That will still hold all of my things while keeping me organized. So what fits, I can put my 12.9" iPad, with case and Magic Keyboard in the spot for a laptop. I can also put my 8x10 spiral bound planner, a notebook, mouse, small cable case, pencil case, water bottle and purse pouch with all my essentials. So my water bottle does not fit inside the side bottle pockets, so I do have to put it inside the bag it's self, but that's a no issue. The shoulder straps seem sturdy enough and the color beige/cream looks nice with my neutral wardrobe. Overall I'm happy with the purchase. We'll see how it travels over the next year as I have quite a bit of traveling and work do to offsite this year. j
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Reviewed in the United States on January 17, 2023
★★★★★ 4
Stylish, High-Quality Backpack. A Great Upgrade from My Usual Black Bags!
Color: Pinkish Grey, Size: 14-Inch
I’m really impressed with this backpack. I usually stick to black bags, but I decided to try something different, and I’m so glad I did. The color is beautiful—unique without being too loud—and adds a nice pop. The quality of the material feels top-notch. It’s sturdy yet lightweight, and the stitching and zippers seem very durable.
Overall, this backpack feels well thought out in both design and function. Highly recommend if you’re looking for something professional, stylish, and practical.
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Reviewed in the United States on June 25, 2025
★★★★★ 5
Buena compra
Color: Apricot, Size: 14-Inch
Calidad, Color, espacio.
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Reviewed in the United States on May 1, 2026
★★★★★ 3
Gorgeous, but some big downsides
Color: Pink-upgrade, Size: 14 Inch
I wanted to like this bag SO BADLY. I’ve owned other backpacks by this brand and they have never disappointed. This one though, was a close miss. The color is absolutely GORGEOUS and I almost kept it for that reason alone. However, two big things made me return it: 1) The inside is no longer padded. If you had purchased older Golf Supags backpacks, the interior had padding to help the backpack keep its shape and not slouch over on the floor. That’s gone, so this one does flop over a little. But the big deal breaker: 2) holy cannoli, these straps are WAY too long! On the shortest length, the backpack is still hanging almost halfway down my back. It looks weird, but it’s also not ergonomically good for your back. The actual strap part (with padding) is so far back that the thin seatbelt adjustable material is the only thing on my shoulders. Super awkward. I’m not necessarily a small person either, so I shudder to think what this would look like on someone more petite than me.
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Reviewed in the United States on July 15, 2025
★★★★★ 1
Flimsy and cheap
Color: Apricot, Size: 14-Inch
This is the second time I bought a purse online... First was on temu ... Expected to be trash and it was... So I bought this one... And idk what's worse, the temu purse or this one.. only buying them in person from now on unless Its 100% obviously sturdy and of good quality. The fabric and entire purse itself is cheap thin fabric .. not only does it feel cheap it looks cheap. I'm not one to write comments but I'd return it if I wasn't so damn lazy... It feels like the play purses your parents would give you as a kid. Not worth the money. I guess it has pockets, but I feel like my things would be sitting on the floor even though my purse is a barrier and feels like my belongings are not protected in the slightest... Especially not a laptop as advertised.
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Reviewed in the United States on April 5, 2025