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Description
Human YTHDF3 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. |
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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 conjugate are sequentially added to microwells pre-coated with a YTH domain family protein 3 (YTHDF3) 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 YTH domain family protein 3 (YTHDF3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human YTH domain family protein 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | YTH domain-containing family protein 3 (YTHDF3) is a protein encoded by the YTHDF3 gene. Diseases associated with YTHDF3 include Wilms Tumor 1. Gene Ontology (GO) annotations associated with this gene include RNA binding and N6-methyladenosine-containing RNA binding. An important homolog of this gene is YTHDF1. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.8 ★★★★★
Based on 471 reviews
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Product Reviews
★★★★★ 5
A stark, brutish "Petulia" with a pistol in its pocket
It's about time this movie got released on DVD.
It's odd that a film could spawn a remake ("Payback"), a glib nod ("Grosse Pointe Blank") and countless homages ("The Limey," among others) and still be as underseen as "Point Blank."
The lack of a disc certainly didn't help its low profile, but of course this is a challenging, idiosyncratic movie, even three decades later. The plot is simple -- a crook is betrayed by his wife and partner and spends the rest of the movie trying to get what he's owed -- but the editing and narrative structure is unusual. What in the world did audiences possibly make of this back when it was first released?
It's a remarkable film, as startling and innovative as Richard Lester's "Petulia," although admittedly it's thematically much less complex.
This edition is excellent, too. Great sound, great picture and a fantastic commentary by director John Boorman and big-time "Point" fan Steven Soderbergh, who laughingly admits to Boorman that he's ripped this movie off more than a few times. Their chat is more technical than gossipy and deals heavily with the editing, the production (the script was only 70 pages long), the studio's concerns about the picture, the actors, violence, surrealism (is it all a dream?) and Boorman's elaborate use of color (the tones of clothing and sets intensify over the course of the film).
I've gotten a lot of good DVD's this year but in terms of content, presentation and extas, this is one of the best.
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Reviewed in the United States on August 1, 2005
★★★★★ 4
One of Marvins better offerings
Like the Killers before marvin was almost destined to play the part of Walker in this fast moving gut wrenching but always realistic thriller brilliantly collaborated by Marvin and Boorman who had no peer in this type of film.For those unenlightened souls who downplay Marvins career this was the one that to my mind surpassed most of his prior efforts with the exception of the Killers which was above par in all respects.Marvin was ahead of the pack in the 60s playing this type of hard nosed no nonsense gangster type, no other actor came close and type casted him to some extent in this type of role which unlike may actors became a positive in his career. Just to show his brilliance as an actor he gave us later comedy roles which produced more acting accolades than that material for which he was better known namely what we see in Point Blank.He carries the whole movie as did all the great actors of that era and many since which in itself is the hallmark of greatness. Marvin was a man who so perfectly personified the parts he was playing that often other actors looked wooden in comparison.He was one of the rare individuals who could take a small co starring role and end up being the star of the movie, no mean feat when you were up against the best in the business at that time and there were plenty in the 60 and 70s.To my mind Point Blank will always be a MARVIN film and this is not to downplay other good workmanlike performances in the film but it will always rank highly in Marvins body of work which is a cut above his contemporaries in the roles for which he was known and appreciated.As for the film it played out in fairly predictable fashion until the final scenes under the golden gate brige which gave a nice twist and left the viewer unsure whether Walker did in fact take his money or simply leave the scene of the set up empty handed. The obvious answer is that he waited till the coast was clear and took his money. It is hard to accept that he did otherwise.In retrospect a movie that stands up 40 odd years later and is just as watchable as it was in 67. No mean feat.
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Reviewed in the United States on May 20, 2010
★★★★★ 5
A forerunner to Dirty Harry and Lee Marvin shining...
If you liked "The Getaway" or "Dirty Harry", then meet their forerunner.
"Point Blank" is explosive, fast-paced, and still the acting is there.
Good acting that is.
Lee Marvin is at his best. Angie Dickinson. in her strong performance, is as beautiful as ever. Keenan Wynn and Carroll O'Connor play their parts to the hilt and it truly shows. John Vernon (who was The Mayor in "Dirty Harry") plays a slimy type with diligence and very believable.
Add the killing pace of the entire picture, and you have a hot item, as sharp and cutting as "Film Noir" can be.
Yes, because this is still a "Film Noir", despite the fact that it was filmed in Color and in the mid-sixties.
John Boorman ("Hell in the Pacific", "Deliverance" and "The Emerald Forest"), skillfully "color coded" the entire movie, bringing it from absolute colors at the beginning, to more red-tinted ones towards the end.
The only difference from a true "Film Noir" is its fast-paced storyline, that would lead us to movies as I have mentioned above.
Marvin's minimalistic acting, but forceful presence, is enough to fill every frame of the movie with tension, action and complete mayhem.
Compare him in "The Dirty Dozen" and "The Big Red One" and you will see what I mean.
A big plus was the release on DVD. An excellent transfer with a sharp picture resolution, a clearcut sound, make it a very enticing experience to watch it at home.
This is not just a Highly Recommended title. It is simply a Must!
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Reviewed in the United States on April 20, 2007
★★★★★ 5
Whiter teeth with out the trays
Size: 3.8 Ounce (Pack of 1)
Love this toothpaste! My teeth are a lot whiter and brighter. Doesn’t leave a bad taste in my mouth. Highly recommend this product.
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Reviewed in the United States on May 24, 2026
★★★★★ 5
smile time!!
Size: 3.8 Ounce (Pack of 1)
Love the whiteness! Works in a few weeks!
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Reviewed in the United States on June 8, 2026