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Description
Human SEMA3B 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 specimens using EDTA or heparin as an anticoagulant and centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-Test Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 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 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 ng/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 semaphorin-3B (SEMA3B) 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 semaphorin-3B (SEMA3B) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Semaphorin-3B ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Semaphorin-3B is a protein encoded by the SEMA3B gene. The semaphorin/collapsin family of molecules plays a key role in guiding growth cones during neuronal development. Secreted proteins encoded by members of this gene family are important in axon guidance and have been shown to function as tumor suppressors by inducing apoptosis. Diseases associated with SEMA3B include phenylcyclic drug abuse and lung cancer. Its relevance also includes neuroscience. | |||||||||||||||||||||||||||||||||
| 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, cell culture supernatant |
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4.1 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 4
Comparable to Major Name Brands in price and quality
Size: Newborn (30 Count)
Good value for the price , comparable in value and quality of major name brands. Visually attractive and detailed product description on packaging. Newborn size package is very small and well packaged for easy storage. Diapers are made to fit comfortably to newborns body and manufactured to wick moisture into the diaper away from babies skin and so not to leak. They absorb a fairly good amount of liquid with no problems. The adhesive tabs stick well and do not come lose.
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Reviewed in the United States on May 31, 2026
★★★★★ 5
Lifesavers for sensitive skin!
Size: Size 6 (108 Count)
These are the only diapers we love after trying countless other brands. We struggled with so many issues, from diapers that felt cheaply made and rough, to constant leaks, or scents that caused horrible breakouts on my son’s sensitive skin. The Mama Bear Gentle Touch diapers are the first ones I have found that are truly soft, completely leakproof for us, and do not cause any irritation. They are lifesavers, and it is a genuine nightmare when we run out. They perform great overnight with no leaks, and they hold up well during active daytime play. We do get a few leaks, but it’s when my son chugs the water. I don’t think any diaper is saving that. The fit is consistent, the tabs are secure but still stretchy, and buying the bulk pack is a good value. If you have a child with very sensitive skin, I highly recommend giving these a try.
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Reviewed in the United States on October 10, 2025
★★★★★ 5
Wish we had these diapers with my first 3
Size: Newborn (120 Count), Size: Newborn (120 Count)
I am 39 been married 22 years and I have 4 kids. We have always had our kids seperated over the years and I have always used Luv's with my first 3 all other diapers leaked badly, never lasted long or gave the kids rashes. Luv's worked a little better on leaks at night none of the these diaper prevented leaks on sheets and bed while sleeping u always had to change there diapers multiple times a c there skin was always wet. With my first 3 kids we did not have the choices we have now days.
Well my 4th child is now 18 months and I had hopes of no more changing sheets in the middle of night. I finally got that with these diapers now I have learned all 3 above brands are garbage. Now they feel like paper to me and a waste of money. When I have ran out of diapers my husband has picked these diapers up at the local store. What happens wet clothes wet beds rashes. I never use these donate them everytime. When I see people say these are cheap diapers and they leak I have no clue what your talking about and my guess is they are buying the wrong size or not putting them on right. Alot of parents don't even realize they are using the wrong sizes for there child and if u are not buying the right size or put them on correct yes u will have leaks. The only time my sons skin is wet is when the diapers have been left on way to long and his peed alot. So if he slept 10 hours over night with no diaper change and he drank alot before bed this happens rarely but when he does this happens. Now while he was tiny no I didn't have this issue. I will say these are amazing diapers and I have tryed many diapers during this time more expensive and less these are the only diapers I will buy. It's also the first time ever with all my kids I didn't wake up many times with newborn having wet clothes and soaking the bed all thoughout the night some night soaking me too. There overnight diapers are really amazing too.
I would recommend these to anyone and I would never do so for luv pamper huggies I see those as a waste of money cheap diapers. These are perfect, know the correct size buy bigger keep diapers above belly button to many buy them to small.
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Reviewed in the United States on July 21, 2024
★★★★★ 3
Frequent leaks
Size: Size 2 (184 Count)
These diapers leak through frequently. They also run small so make sure to size up. My LO wears a size 1 in other diaper brands but mamabear’s size 1 was tiny. Had to exchange for a size 2.
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Reviewed in the United States on June 2, 2026
★★★★★ 5
AMAZING quality & price
Size: Size 7 (80 Count)
Great diapers at an unbeatable price—by far the cheapest option I’ve found. They’re also the only brand that fits my toddlers well since they aren’t potty trained, which has been a huge plus.
The quality is solid for the price, and they hold up well overall. The only small downside is that the fit around the legs can occasionally shift a bit.
Doesn’t irritate the skin often. Great moisture absorption.
Overall, they’re well worth it and a great budget-friendly option.
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Reviewed in the United States on May 13, 2026
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