Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 8 - Jul 13
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Mouse TIGIT 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: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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. 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, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against T-Cell Immunoreceptor With Ig and ITIM Domains Protein (TIGIT). 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 T-Cell Immunoreceptor With Ig and ITIM Domains Protein (TIGIT) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse T-Cell Immunoreceptor With Ig And ITIM Domains Protein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | The Ig and ITIM domain protein T-cell immunoreceptor (TIGIT) is an immunoreceptor present on some T cells and natural killer (NK) cells. It is also known as WUCAM and Vstm3. It binds with high affinity to CD155 (PVR) on dendritic cells (DCs) and macrophages, and with low affinity to CD112 (PVRL2). | |||||||||||||||||||||||||||||||||
| 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, 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
4.8 ★★★★★
Based on 1149 reviews
Sort
Product Reviews
★★★★★ 5
Simple and streamlined to meet anyone's needs
Format: Paperback
This book rocks! It provide a simple and structured way to define, develop and deliver a "Kick Ass" presentation that you can project or just use from the podium. It bring the audio and visual together to compliment a story to be shared. It has taken the hassel factor out of designing a presentation for anyone on any subject. Thank you!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 11, 2014
★★★★★ 2
Lasting value but degraded presentation
Format: Paperback
I've been a BBP fan since the first edition in 2005. The second edition in 2007 was a significant update and refinement. The latest, 2011 edition, is not worth buying if you already have the second edition -- the information is essentially the same but the 2011 is printed on cheap paper and the contrast between text and background for most of the illustrations is lousy, e.g. black text on a dark grey background, white text on a light grey background. I found it very difficult to read. The online version (O'Reilly Safari Books OnLine) is slightly better because it allows you to magnify the low contrast images which makes them slightly more legible. Thankfully, access to the online version is included with the ink-on-paper book free via a scratch-off, lottery ticket-type insert at the back of the book. If I had to do it again, I'd skip the paper book from Amazon and just purchase electronic access via O'Reilly. This version is more legible and you can print important pages that you might want hard copies of to highlight, mark-up, etc.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 18, 2011
★★★★★ 4
Powerful for the presenter
Format: Paperback
My boss's boss recently told me that our organization's presentations are boring data dumps that make her wish she was somewhere other than there listening but not really hearing or caring about this slide and dreading the next. She suggested that since I was going to be doing a lot of presentations in the future I should avoid that method and do better presentations. I immediately sought advice from our best presenter and learned that his method came from his gut feelings about what worked better. He has good instincts but to codify what he explained I decided to buy a book to learn why his approach works. The closest book with an effective explanation I could find is "beyond bullet points". It captured and expanded on the instinct my new mentor explained so that I could now understand how to use the slides to supplement the story and how to interweave the data so the whole presentation is more meaningful and memorable. I recommend this book to anyone looking for more of an answer on how to improve their presentations and eliminate the data dump in bullet points.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 17, 2014
★★★★★ 5
Good purchase and superb seller.
Format: Paperback
This is a very good book. I was very satisfied with the service I received, I got the book before the expected time. I was really supprised with the shape of the book...it was a used book but looked like brand new. I say this was superb purchase and the seller was top notch.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 4, 2013
★★★★★ 5
Great for Presentation Education
Format: Paperback
It has been some time since I have utilized Beyond Bullet Points, but this is one text book that kept with me. The point of the book is to help a presenter get their point across to the audience through Powerpoint presentations, and the book does this well. I am sure there are editions other than this one at this point, and I would definitely recommend looking into them in order to keep up to date. Starting with this book is a great idea due to its simplicity.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 24, 2014
recommand products
Centric 12-14 Ford F-150 C-TEK Rear Rotor - 121.65133
167.67
Mishimoto 99-06 Chevrolet Silverado 1500 V8 Blue Silicone Hose Kit - MMHOSE-RADO-99BL
125.47
WeatherTech 2017+ Jeep Compass Front FloorLiner - Black - 4412051
126.95
xTune 10-13 Chevy Camaro CCFL Halo Projector Headlights - Chrome (PRO-JH-CCAM10-LED-C) - 9040504
222.04
Borla Universal 3in Stainless Steel 3pc V-Band Clamp w/ Male and Female Flanges - 18009
87.99