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Description
Human PDCD1 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes and remove the supernatant for analysis. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove 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 250 pg/mL). Then dilute to the following concentrations: 250 pg/mL, 125 pg/mL, 62.5 pg/mL, 31.25 pg/mL, 15.625 pg/mL, 7.8125 pg/mL, 3.90625 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 250pg/mL standard working solution into the first EP tube and mix thoroughly to make a 125pg/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 a Programmed Cell Death Protein 1 (PDCD1) 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 Programmed Cell Death Protein 1 (PDCD1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Programmed Cell Death Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Programmed cell death protein 1, also known as PDCD1 and CD279 (cluster of differentiation 279), is a protein on the surface of T and B cells that regulates the immune system's response to human cells by inhibiting the inflammatory activity of T cells. Expression of PD-L1 on tumor cells inhibits anti-tumor activity by engaging PD-1 on effector T cells. PD-L1 expression on tumors is associated with decreased survival in esophageal, pancreatic, and other cancer types, highlighting this pathway as a target for immunotherapy. | |||||||||||||||||||||||||||||||||
| 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 | 3.9-250pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell culture supernatants, and other biological fluids |
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4.4 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 5
Definitely worth a read
Format: Kindle
This book was everything. It was heart-wrentching, funny, thrilling, sweet, everything. You'll be angry. You'll be sad. You'll be shocked. So much happens and so much is still yet to be explained. I loved it. From the beginning, I was interested in Emmy as a character. She's extremely relatable. She's realistic. She has reactions and feelings that make sense! The boys will make you want to throw things sometimes, but they'll worm their way into your heart eventually. And that ending! Yes! Loved it
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 9, 2021
★★★★★ 5
Kiss
Format: Kindle
Was a great read. What a fantastic Hidden Gem I hope doesn't stay hidden. This was a great darkest reverse harem. The FMC was kidnapped in abuse in her past her and her mom are staying on the property of a different Mafia that uses Greek names. Hiding there for safety. The only person that knows is the leader's wife who is friends with the fmc's mom from high school.
The FMC gets involved with the leader son and his two friends. They're all written really well Aries definitely stands out he's a really well-written great character. But I like them all. There is some other woman drama, so heads up there. But I would call this like a medium burn. They get together this book and there are some spicy steamy scenes. But I'm really here for the plot the plot is great the characters are great. I kind of had a feeling of one of the thing is going down from the title of the next book and just a feeling that I got. There are multiple povs it's more the fmc's point of view but we got the guys povs only need them which I really liked.
I really love the story it's really enjoyable I'm so excited for next book and I'm really excited to see that the authors would be putting out a book every month after this book I can say I'm excited to read anything that she writes I'm hoping they're all RH.
There were some like typos spelling errors but it did not hinder my reading experience. Just Small Things having someone do a once-over on could pick up. But honestly like it's is a five-star read in an extremely impressive debut and even if it wasn't a debut I would say it's impressive. It's hard to balance a story with this type of feel a lot of times I think it came out really well and I'm completely enthralled I can't wait for more highly recommend
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 3, 2022
★★★★★ 4
Run little rabbit
Format: Kindle
Though a familiar trope, it had these teases of intrigue that are enough to keep the story interesting to pull the reader to keep reading, to invest in the plot. Enemies (bullies)-to- lovers, mafia, and RH tropes are all so overworked that there is very little new in the way of a storyline. While I can't say this story is utterly unique, I found myself enjoying the characters and their dynamics together. I look forward to seeing how things go from here between the guys and Emmy, and even how Paloma will fair.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 14, 2022
★★★★★ 5
awesome
Format: Kindle
I’ve never read a book by this author before, but she knocked it out of the park! Thanks so much for thinking of us readers who are left with book hangovers and just need the next book in the series. I just downloaded the next one. I don’t normally write reviews but this author needs to be out there so everyone can find her. Great book, excellent characters and pacing. Wonderful storyline.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 18, 2022
★★★★★ 5
pleasantly surprised!
Format: Kindle
So, I’ve been on the Find Me (WITSEC) book train and trying to find a book to hold me over until the second book comes out and omg now I’m going to have to do the same for THIS book!
I was drawn in from the beginning. The characters, the multi POV (which is one thing WITSEC lacks), the world building! Excellent! I love a good grovel and I think these guys are headed that way now! I can’t wait to see where the next book takes these characters!
Now….what do I do while I wait?
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 29, 2021
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