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Description
Human EIF4G1 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 capture antibody against eukaryotic translation initiation factor 4G1 (EIF4G1). 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 eukaryotic translation initiation factor 4G1 (EIF4G1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Eukaryotic translation initiation factor 4G1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Eukaryotic translation initiation factor 4G1 (EIF4G1) is a protein encoded by the EIF4G1 gene. The protein encoded by this gene is a component of the protein complex eIF4F, involved in mRNA cap recognition, ATP-dependent unwinding of the 5'-end secondary structure, and mRNA recruitment to the ribosome. EIF4G acts as a scaffold, interacting with mRNA and other components of the EIF4F complex, as well as PABP and EIF3, to produce five transcript variants encoding four different isoforms. It has been shown to interact with MKNK1, EIF4A1, EIF4E, MKNK2, and PABPC1. | |||||||||||||||||||||||||||||||||
| 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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4.6 ★★★★★
Based on 12 reviews
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Product Reviews
★★★★★ 5
Battle tested
I have re-purchase this product a few times. It is great for razor bumps and clogged pores.
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Reviewed in the United States on May 28, 2026
★★★★★ 5
Great Item to get!
Thank you. Great product!
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Reviewed in the United States on May 24, 2026
★★★★★ 5
Ruthless, nail-biting action thriller from "Exorcist" director William Friedkin.
BOTTOM LINE: If you enjoy tension & suspense then you'll be hard-pressed to find a better experience than SORCERER. I loved it and feel it has a high replay value that offsets the discouraging lack of bonus features. 5 STARS
THE STORY: Four shady criminals from various countries are on the run from trouble. Each eventually ends up in the desolate backwards village of Vera Cruz, deep in the South American jungle. Arriving there illegally, none of the strangers possess proper documentation with which to leave once the heat is off. They are trapped, and the only work in the village is at a nearby oil drilling site. When saboteurs blow the rig up the oil company must quickly put it out & recap the well, to maintain its profitability. The explosives needed to blow out the raging fire are stored at the company headquarters - some 200 miles away. Complicating matters further, the TNT has become dangerously unstable due to improper storage conditions, so it cannot be flown to the site but must instead be delivered overland by truck. Knowing the danger, the company agrees to pay $10,000 each for drivers willing to tackle the high risk job. Desperate to earn the much-needed money with which to buy their way to freedom, all four criminals volunteer. Though in essence a suicide mission, each man has no choice if he hopes to leave the village and get on with his life. Who will make it?
THOUGHTS: Released a week after the first STAR WARS, and sadly lost in the shuffle, SORCERER is a terrific film. Maybe director William Friedkin's very best. Yes, it is a thinly-veiled re-working of the classic French thriller THE WAGES OF FEAR but don't let that deter you. In many ways Friedkin's adaption is superior. JAWS alumni Roy Scheider is terrific here as a seedy driver for a mob hit gone terribly wrong and the other actors are equally adept in their roles. SORCERER is beautifully photographed and tightly edited for maximum tension. Completely done practically and on actual locations, (being shot back before the advent of CGI and wirework), the action set pieces are among THE MOST intense & nerve-wracking I have EVER seen on film. Period. The rope bridge scene? Two words: Holy crap. I was literally on the edge of my seat throughout the entire sequence, my heart thumping and my hands clutching the arms of my chair in a death grip. If this scene doesn't make you break out in a cold sweat and get your pulse pounding then you might want to check with your doctor because you're probably dead. This sequence alone makes the purchase of SORCERER completely justified. I feel the first half of the film could have been tightened up some, as it spends the first 45 minutes or so setting everything up, but taking it for what it is it's worth the wait because the last half will have you gasping in disbelief, jumping in your seat and chewing your nails right down to the first knuckle. Simply great stuff. I only regret that I hadn't discovered SORCERER before now, but what a fantastic find.
THE BLU-RAY: In a word... magnificent. The remaster for SORCERER is an audio & visual triumph. This is THE version to own. While sadly lacking in extras, the hi-def film transfer looks so impressive that I am more than willing to overlook the bonus shortcomings. The Blu comes packaged in a sort of mini book binding that sports a brief excerpt from William Friedkin's autobiography, detailing his work on the film and his feelings about its new lease on life with this Blu-ray. I've never been much of a fan of the director himself because he's always come across like a snobby, pompous, egotistical ass. Surprisingly, he acknowledges this fault and describes how it lead to his eventual downfall in Hollywood. But I cannot deny that he's made a few really good films during his career.
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Reviewed in the United States on January 30, 2016
★★★★★ 5
A Perfect Film
It has been a long time since I have been so surprised by a film; I can remember hearing about it when I was a kid, but like so many others at the time, 'Star Wars' was my childhood obsession. That a being said, I also have to say that this movie is generally timeless; with the exception of the cars and the music sometimes(still an excellent score by Tangerine Dream...), there is nothing that falls to the side as 'dated'. The country the 4 men flee to is just miserable, and probably looks the same now as it did then...just behind the times in so many ways.
The set up of the 4 men starts things off with bang--literally. The scenarios are very different for each individual, but each equally satisfying. There is an excellent church robbery scenario that rounds things off with Roy Scheider, which includes a trademark Friedkin vehicle sequence(which--like many other scenes in this film--comes with a surprising jolt and a direction you didn't see coming...). Each backdrop story could pretty much provide for a film of it's own. Scheider is excellent, and this film reminds me why I was always a big fan of his. That being said, the other acting is all just right on the money as well; you can feel both their desperation and sweat, and sympathize with every stumbling block they encounter along the way. From the corrupt cops to the bandits they meet on the road(a favorite scene of mine in any movie...), you are thrown generally everything in the book as far as a crime/thriller movie go. And in this case it is all successful. There were more than a dozen scenes that hold up to or outshine anything that has been put out in the last 10 years to me. It is a reminder of the skill behind this project across the board, and the booklet with the blu-ray is really enjoyable to read, especially after seeing the film, as it fills in some curious questions about how certain things were done, and the difficulties the production encountered.
The look of the film is breathtaking at times; all of the camera shots of the landscapes, kids, animals, elderly soda vendors, and crowded village streets are very candid and realistic. While the plot is somewhat simple, the intensity and driving force behind the 4 men is unforgettable; and while they are all different in their ideals, beliefs, and character, they all manage to somehow come together by fate to carry out the one thing that could save them from a life of poverty or consequence by law. And everyone meets some kind of fate you don't see coming.
I have always loved Friedkin's work, especially when he delves into the darker side of mankind; However, this is the 1st movie in quite some time that has jumped into my top 5 favorite movies. It again reminds me how people w/talent to burn make timeless work, period. I am so happy that Friedkin was able to get this released as he saw fit, and it truly is a gift to those who like him, the genre, the actors, and the feel of such a film...
There is humor and philosophy throughout the film here and there as well; the screenplay is top notch, and the dialog is fascinating; it is one of the films you watch where you are never reminded it's a film. You get wrapped up in it, and do a lot of replaying certain sequences over and over. While the 'bridge scene' is the most famously known sequence in the film, there are countless others that are certainly equal, although the bridge scene is a pure nail biter(not to mention technical feat, as further explained in the booklet...).
The use of sound is also pretty incredible, especially coupled with the visuals; from the sounds of several surprise explosions, to the sound of the knife blade being flipped out, this film uses sound as a performance of it's own; it supplies a huge/equal part of the overall canvass and to feeling like you are there in particular a situation, not to mention hearing things as they actually probably would be heard; the sound is never overdone for the situation, and always effective. The creaking wood and rushing water of the bridge scene is a great example. Also, while the dialog is considerably less than in most movies, it is always very well written, realistic, and just another positive strength; but even when there is no dialog, and just simply sound and the expressions on the characters faces, it more than serves the point trying to be made. The character hit-man Nilo has very few lines, yet his character and performance are equally strong as anyone's here. The small visual details work in the same way the sounds ones do, such as the soldier accepting a bribe by scraping the money to himself with a fly swatter, the bar tender wiping off Roy Schieder's silverwear on his own shirt before passing them to him, to the cop popping open a Coke bottle with the cocked barrel of his gun...all small examples of going that little extra bit to make something potentially average something more. And these types of moments and details are throughout the film. A favorite to me is in the church scene in the beginning of the film where the priest mentions to the happy couple something to the effect of 'not being obsessed with material things', and you can see the groom roll his eyes, all while his bride with the black eye stands next to him(they show a close-up of her when the priest say he hopes their marriage is filled with blessings, but she still didn't look too happy to be there...).
Mr. Friedkin wanted this to be his 'magnum opus', and I think he well succeeded. It really does still outshine so many films and the people involved that exist nowadays; Like Stanley Kubrick and John Huston, I can see the obsessive side Friedkin had to create something unforgettable no matter how much time or money it took; it is quite a risk, and sadly one that doesn't always pay off; but it is an effort that is well appreciated, and something not as many filmmakers take on nowadays.
As far as it being considered a remake, I would have to say this is no more a remake than something like Herzog's 'Bad Lieutenant: Port of Call, New Orleans' was of the original 'Bad Lieutenant' film; some of the story elements are similar, but a lot of those elements are taken to new and different places with all the people involved with the production; and while 'Sorcerer' gets it's story foundation from both a novel and a film, you can see them much more as an influence, and not the basis. It is more the taking of a loose idea and making it to all your own, not an imitation of it.
I have watched this 6 times since it came out, and it still seems to expose a new layer or understanding each time, not to mention the brilliance of so many scenes that capture the moment spot-on. This film is pure genius to me, and the surprise release on blu-ray has made my month. It is a piece of work that warrants all the 5 star reviews it has gotten, and Mr. Friedkin should be more than proud of this piece of work.
I hope it reaches the full audience it deserves now, esp in the blu-ray format. I think it will.
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Reviewed in the United States on April 30, 2014
★★★★★ 5
The finest hour of William Friedkin
William Friedkin was undoubtedly one of the 1970s greatest and most influential directors. He only made 3 pictures during the decade, but all are landmark films. The first was The French Connection (1971), the film that won him his best director Oscar & the film won best picture. His next film, The Exorcist (1973) is probably his most famous/infamous & and it’s the only movie that’s still a truly terrifying experience, something to endure as much as to watch almost lol. Sorcerer (1977) was his 3rd and final film of the decade, and in spite of or perhaps even because of how it was a largely forgotten film for so long (after seeing it on cable nearly 30 years ago, it was a movie I simply could not track down until the release of this Blu-Ray) I really think in so many ways it ranks right along aside his first two 70s classics, and perhaps even surpasses those films in terms of pure audacity. The rope bridge scene is as intense as a set piece gets and is worth the price of admission all on its own, but the film as a whole is a clinic in lean, economical filmmaking. And the score by Tangerine Dream kicks serious a**. It might be because of his 70s trifecta Sorcerer is the only one that is still almost sort of like a “new film” too me, as I’ve only watched it maybe 3-4 times and have seen the other 2 dozens of times, but for my money Sorcerer is master filmmaker William Friedkin at the absolute peak of his powers.
MD2020 gives “Sorcerer” 5 out of 5 ⭐️s and says it’s a flawless action film and Friedkins best work.
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Reviewed in the United States on January 25, 2024
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