Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Aug 30 - Sep 4
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Human WNT5A 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 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. |
|||||||||||||||||||||||||||||||||
| 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 Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Wingless Type MMTV Integration Site Family, Member 5A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Wingless-type MMTV integration site family member 5A, also known as Wnt-5a protein, is encoded by the WNT5A gene. Disorders associated with WNT5A include Robinaud syndrome, autosomal dominant 1, and autosomal dominant Robinaud syndrome. Pathways associated with this gene include the canonical Wnt pathway and CDK-mediated phosphorylation and clearance of Cdc6. Gene ontology annotations associated with this gene include DNA-bound transcription factor activity and protein domain-specific binding. An important homolog of this gene is WNT5B. | |||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| 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, cell culture supernatants, and other biological fluids |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.6 ★★★★★
Based on 23 reviews
Sort
Product Reviews
★★★★★ 5
Nice
Size: 9, Color: Black
Very comfortable. Worth the money
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 1, 2026
★★★★★ 5
Good purchase
Size: 9.5, Color: Black
Value and quality was good
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 16, 2026
★★★★★ 5
The "Lazy Professional" Look: Is Hands-Free Luxury Actually Real?
Size: 9.5, Color: Black Napa Leather
Living down here in Florida, my footwear needs are pretty specific. It’s hot, it’s humid, and I spent half my life rushing from the car into work or a meeting. I’ve reached that age where I value efficiency just as much as style—maybe more. I’ve been eyeing the Marc Joseph New York Hands-Free Slip-on Penny Loafers for a while, and after putting them through the wringer, here is the honest truth from someone who just wants to look sharp without the hassle.
The "Just Step-In" Reality
Look, the big selling point here is the "Hands-Free" tech. We’ve all seen the commercials for those athletic slip-ins, but finding that in a legitimate leather penny loafer is a different game. Does it work? Yes, surprisingly well. The heel counter is firm enough that it doesn’t collapse when you slide your foot in, but it doesn’t feel like a piece of plastic digging into your Achilles once you’re in. For those of us who are tired of bending over or hunting for a shoehorn every morning, this is a genuine quality-of-life upgrade.
The Florida Factor: Comfort and Style
The leather is actual calfskin (on most models), which is a must for the Florida heat. Synthetic shoes turn into a sauna within ten minutes, but these breathe reasonably well. The aesthetic is classic—it’s a "professor" shoe through and through. You can wear them with chinos and a blazer for work or throw them on with some nice jeans for a weekend lunch.
Inside, they’ve got a gel heel insert and a padded footbed. It’s not quite "walking on a cloud"—let’s not over-hype it—but it’s a massive step up from the hard, flat soles of traditional dress loafers. I’ve spent four hours on my feet lecturing, and my arches didn’t hate me by the end of the day.
The Sizing Gamble
Here’s where you need to be careful. The consensus from other guys (and my own experience) is that the sizing is a bit of a coin toss. They tend to run a little large and sometimes wide. If you have narrow feet, you might find the sides "gaping" or flaring out when you walk, which kills the sleek look. I’d recommend ordering a half-size down if you’re usually between sizes.
The Breakdown
The Pros:
True Hands-Free: You can actually put these on while holding a coffee and a briefcase. No hands needed.
Legit Materials: The calf leather feels premium and smells like the real deal.
Versatility: Perfectly bridges the gap between a "car shoe" and a formal loafer.
Comfort: The gel insert makes a noticeable difference for long-duration wear.
The Cons:
Sizing Inconsistency: They can run wide, leading to some "heel slip" if you don't get the perfect fit.
Break-in Period: The heel is stiff by design (to allow for the slip-on feature), so expect a day or two of minor stiffness.
Price Point: They aren't cheap, though often on sale.
Value for Money
Are they worth it? If you’re comparing them to high-end Italian brands that cost $500, these are an absolute steal. They look 90% as good for a fraction of the price. However, if you're used to $60 mall shoes, the jump to $150+ might feel steep until you realize you’re paying for the convenience of never having to touch your shoes to put them on. For a daily driver in a professional setting, the value is definitely there.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 10, 2026
★★★★★ 5
Step In Look Good
Size: 11.5, Color: Cognac Napa Leather
I am wearing these shoes as I write this review, and they look and feel great. I have a bit of a disability with drop foot on the right leg and bilateral neuropathy and have been wearing step in shoes for about 10 years. But the first company that introduced leather dress and casual shoes stopped making them and now only makes sport and casual shoes. So I have been searching for some that meet my need for shoes like this and this company has given me three pairs, so far. Easy to get into, comfortable to wear, and good looking. And they take polish very well, too. (Remember how to do that?). We may buy another pair or two in different styles as well.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 26, 2026
★★★★★ 4
Runs tight. Skinny and smaller than other brands.
Size: 8.5, Color: Brown
Comfy and lots of support. Cushiony. Beware: the brown I ordered runs small. Tighter than other brands.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 23, 2026