Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 23 - Jul 28
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Mouse 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. Preparation before testing: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 10ng/mL). Then dilute to the following concentrations: 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.3125ng/mL, 0.15625ng/mL, and 0ng/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 peroxidase (HRP) catalysis 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 | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Wingless Type MMTV Integration Site Family; Member 5A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| 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
4.2 ★★★★★
Based on 7 reviews
Sort
Product Reviews
★★★★★ 5
Totally engaging story!
Format: Kindle
Here’s the thing about writing reviews. I sometimes feel that I keep writing the same thing. But what I try to express is whether I enjoyed the story. If I felt the characters had chemistry. If the dialogue and narrative held my interest. If the scenes flowed and moved the story on at a good pace. If not, I typically make notes along the way. When I finished this story, as I have with a few other books, I found that I had not made one comment. You know why? Because I enjoyed the characters, the dialogue, the narrative and the storyline. No spoilers from me, remember. But I will sneak in a comment about the sex scenes—hot! I loved the power dynamic and play scenes. I also need a day or so to recover from a story that held me hostage, in a good way. Just sayin’.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 4, 2024
★★★★★ 5
a story with fire
Format: Kindle
The story has a great balance of spice and story. The characters are well developed. It has great ups and down throughout. Well worth the read.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 22, 2025
★★★★★ 5
Oh My Anna Has Done It AGAIN
Format: Kindle
I have been a hard core Anna Stone fan from the first book I read by her...Behind Closed Doors....then I picked up Being Hers the first of the Irresistible Bound series! I fell in love with Vanessa and Mel so deeply I couldn't stop falling deeper in the world that Anna created with her Liliths Den ladies! I devoured her next Mistresses Series even more so then the first. When she told me she was moving to LA with a new set of Ladies I was excited to see what she was going to do and I have to say Club Velvet is even more alluring then Liliths Den and this new set of Ladies I love them all already!!!
Simone is even more alluring then Camilla was for me and I fell hard for Camilla! I love that she was straight to protective with Jade from the start but not overly so. Simone is sweet and demanding and just perfection rolled into the perfect package and just what Jade needed! Both were afraid of falling and giving up that trust and vulnerability of falling in love because of past wrongs and being shown how devastating it can be when its given to the wrong person. The way they handled it just broke my heart for the slight misunderstanding that could have been avoided by trusting each other. But once again Anna has brought fourth just the right conflict for our ladies to get through while also building their characters to a level of complete understanding that only she can do.
I love Jade and Simone and can't wait for hopefully Valerie with her Nanny next **fingers crossed** and hopefully even a wonderfully meet up with Vanessa and Mel! Both Olivia and Elle also excite me to learn more of them!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 25, 2024
★★★★★ 3
Writing was decent, characters were decent, the plot was decent, and the spice was excellent
Format: Kindle
TL;DR: 3 stars, writing was decent, characters were decent, the plot was decent, and the spice was excellent.
This is the second book I have read by Stone. I thought it was pretty mediocre with everything except the spice.
The writing was okay, it wasn’t terribly descriptive or informative. I really hate how Stone constantly has the characters ask questions in their heads after a conversation. I don’t think that is a very good writing mechanism, and she does it frequently.
The characters were fine. I don’t think they developed all that well. Jade seemed to work through her issues with her predatory professor, but it still seemed like she was a bit harsh afterward. Simone seemed to be pretty much the same all the way through. She was just less restrained at the end.
The plot was fine, it wasn’t anything to call home about. It just seemed like a poorly developed mechanism to get the MCs together. The third act breakup was terrible. If you think someone is the love of your life, and they do something wrong that upsets you, then you don’t just run away. You work on it with them if they are willing. Simone was extremely apologetic and wanted to work through it was Jade, but Jade walked away. I honestly think that Jade was the one that should have tried to mend it because she was the one who walked away after Simone tried to work it out.
The spice was very good in my opinion. I love a book with positive BDSM material. This book did not disappoint in that regard. The aftercare was also phenomenal.
Overall, I would recommend this book if you want spice with a low plot line and character development.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 5, 2024
★★★★★ 4
Not a bad story
Format: Kindle
I liked it. It was of course a lesbian romance, which I have no issues with. I thought for the most part the story was well written. One problem I did have was I can't understand why Jade can tell Simone about her past with Phillippa, but has a problem telling Simone she's going to meet her to basically tell Phillippa off. That to me doesn't make much sense.
I mean she could have told Simone she was meeting her and that it was something she had to do by herself so she could move on. I just had a problem with it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 14, 2024