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Description
Mouse HCAM 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: 1. 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: 1. 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. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. 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. |
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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 enzyme conjugate are sequentially added to microwells pre-coated with Homing Associated Cell Adhesion Molecule (HCAM) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Homing Associated Cell Adhesion Molecule (HCAM) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Homing Associated Cell Adhesion Molecule ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Leukocyte differentiation antigen 44, also known as CD44 antigen, is a cell surface glycoprotein involved in cell-cell interactions, cell adhesion, and migration. The CD44 antigen is encoded by the CD44 gene on chromosome 11. CD44 has been referred to as HCAM (homing cell adhesion molecule), Pgp-1 (phagocytic glycoprotein-1), Hermes antigen, lymphocyte homing receptor, ECM-III, and HUTCH-1. CD44 is expressed by a wide range of mammalian cell types. The standard isoform, consisting of exons 1-5 and 16-20, is expressed by most cell types and is designated CD44s. Splice variants of CD44 containing alternative exons are designated CD44v. Some epithelial cells also express a larger isoform (CD44E), which includes exons v8-10. CD44 participates in a variety of cellular functions, including lymphocyte activation, recirculation and homing, hematopoiesis, and tumor metastasis. | |||||||||||||||||||||||||||||||||
| 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, tissue homogenates and other biological fluids |
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4.3 ★★★★★
Based on 1371 reviews
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Product Reviews
★★★★★ 5
Asurion is AWESOME!
Ok, so let me start with the fact that you should NOT waste your time calling Amazon's customer service if you think you have a claim under Asurion's monthly protection plan. CALL ASURION!
I wasted almost an hour with Amazon representatives that have absolutely no training in how the Asurion plan works, or even any idea if my item was covered. In fact, they told me the mechanized chair I purchased under the plan wasn't covered!
Finally, my call got escalated to a manager who was just as clueless about my coverage, but had the best idea ever... call Asurion!
The smartest thing I did that morning was hang up with Amazon, and reach out to Asurion as I should have done in the first place.
Dealing with Asurion's customer service, after my Amazon call, was like being elevated to VIP status! 😄
Asurion's rep nicely took all my info, including the chair's purchase date and order number. Then they asked that I upload a couple of pictures of the product, along with a written description of the malfunction I had mentioned over the phone.
This is where it gets really good. They told me they'd review the claim to determine if they'd send a repair
specialist or reimburse me the cost of the item. In less than 48 hours, they determined that a payout was in order, and sent me an email that included an Amazon gift card for the total cost of the item minus the taxes I had paid, which I loaded directly into my Amazon wallet. Apparently, Asurion does not include the purchase taxes in their coverage.
But, I can live with that.
All in all, Asurion's claims division get a double thumbs up 👍🏽👍🏽, and the company gets big ups from me for not only standing behind their protection plan, but doing so expeditiously!
Special note: If you buy a lot of stuff from Amazon, and have Asurion's monthly protection plan, scroll the item listing before pulling the trigger on your purchase... you need to see the green streamer that indicates your product is protected by the plan. If you don't see that, check to see if there are alternative products that might be covered. There are very few electronic or high ticket items that aren't covered, but looking for that green banner in the item description that says it's covered is key to foregoing any future headaches.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 26, 2026
★★★★★ 5
If they can't fix it they will replace it.
This protection plan that i pay monthly for paid for itself ten fold. My 3D printer just quit working and after the manufacturer warrenty had ran out at that. I submitted my claim they emailed me a shipping label in which I shipped out my printer that same day. Two days later they received my printer determined it couldc not be fixed and sent me a Amazon gift card for the full price of my printer. Today I will be receiving my new 3d printer thanks to Complete Protect. I highly recommend anyone and everyone to purchase this policy. This warrenty is like having a home warrenty like Homeshield but for your electronics and probably other stuff that i just have not read to see what else. This insurance will save me so much money over the corse of this year and every year after that. Thank you Complete Protect.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 22, 2026
★★★★★ 5
"Outstanding support and a seamless process!"
Great experience with Asurion insurance through Amazon. I wasn't sure how to register my items for the monthly plan, but the customer support team in the chat was incredibly efficient and guided me through the entire paperwork process smoothly. Furthermore, the service I received at the repair shop (Break Fix) was excellent. The whole process was seamless, and I received my digital gift card without any issues, which I’ve already applied to my Amazon account. I am very grateful for their professionalism and how easy they made everything. Highly recommended!"
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Reviewed in the United States on June 10, 2026
★★★★★ 5
Received giftcard money to replace my plan-protected malfunctioning motherboard
I've been subscribing to the Asurion protection plan for a while. Recently, an Asus Prime PC motherboard I had bought a few months ago on Amazon stopped working well. I ended up considering filing a claim with Asurion to see if they would either fix/replace it or issue the purchase price money, so I could buy the failed electronics replacement component again for my custom-built PC. I went to the protection plan's website and started the claim filing process but couldn't find the category my failed component would be under in order to select it from the drop-down list. I ended up messaging the live customer service. The chat messaging was somewhat slow, the agent told me they were experiencing some communication delays throughout their systems. Ultimately, the agent was very kind and understanding, and completed the claim filing process for me on her side by asking me to provide the necessary info such as the item's product name, date of purchase, price before taxes, etc. I received a printable UPS shipping label in the email, printed it, and returned the item in the mail. Approximately 2 days later, I received an Amazon giftcard code in the email with the dollar amount equal to the item's price before the sales tax. Then I redeemed the giftcard code on the Amazon page and proceeded to apply the giftcard money to my next purchase of a replacement Asus prime motherboard, and it all worked out successfully. Therefore, I had no issues with the protection plan service and recommend it to others.
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Reviewed in the United States on May 24, 2026
★★★★★ 4
Fast and easy but I dont trust the process to be fully automated.
The process was very smooth and fairly quick. The only challenge I had was that the CLAIM status showed complete but I had to manually go look up the status. There was no email or notification it was closed. After manually checking the status, I called customer support and they said since I called in, they would process the payout. I wonder if I didnt call in if they would have processed it? Also, the phone rep said they would issue me a amazon egift card but then the next day I received a email saying they mailed out the payout. Very poor communication but so far the process was easy and fast.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 1, 2026
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