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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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4.7 ★★★★★
Based on 24 reviews
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Product Reviews
★★★★★ 5
Healthy and delicious
Flavor Name: Variety Pack, Size: 12 Fl Oz (Pack of 12)
Let’s compare to Alani! I was a HUGE Alani consumer, wanted something healthier, tried more energy drinks than I can count until I ended up HERE with Bloom! I am very happy about these :)
These have 180mg of green tea caffeine, Alani has 200mg. After switching from Alani to less mg caffeine energy drinks; I cannot handle an Alani anymore! They give me anxiety, and make me feel really bad.
Flavor wise!! Delicious. But let’s compare to an Alani.
Bloom peach mango, in comparison to a juicy peach Alani is a much milder sweetness and doesn’t come across too overwhelming while still being just as delicious, and in my opinion they taste even better!
Bloom cherry lime, in comparison to Alani cherry twist is a HUGE difference. Alani’s leaves a weird chemical after taste that I cannot get past and makes me refuse to drink them. I am a loverrrr of cherry lime drinks, and this one does have a slight medicinal taste, but it’s more like a sweet tart in comparison. It’s still a win to me!
Bloom strawberry watermelon is comparable to Alani watermelon wave!! But not as tart/sour. I am going to be honest in saying this flavor on both brands is probably my favorite! They both remind me of a jolly rancher. Again, I am much happier enjoying the bloom version.
Bloom raspberry lemon is not actually comparable to any Alani in my opinion, I would reach to say Alani pink slush, ONLY because they both have a different sweetness that comes with a “pink” brand drink. I used to love pink slush until the flavor became nothing but carbonation to me.. I can’t be sure but feel the ingredients were changed somewhere along the way, as Alani tends to do. But I really enjoy the flavor of raspberry lemon Bloom, and you can definitely taste BOTH flavors.
Alani is zero sugar, B12, B6 and biotin, 200mg caffeine and artificial sweeteners. After drinking them for maybe three years, I got burnt out.
Bloom has zero sugar, 180mg caffeine, B6, B12, apple cider vinegar, prebiotic fiber, no artificial flavors or color or aspartame.
You can decide for yourself, but this is a side by side comparison:)
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Reviewed in the United States on October 14, 2025
★★★★★ 5
Best energy drink
Flavor Name: Variety Pack, Size: 12 Fl Oz (Pack of 12)
Nice that it comes with three different flavors. They are all great tasting. Great value !
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Reviewed in the United States on April 27, 2026
★★★★★ 5
Variety
Flavor Name: Variety Pack, Size: 12 Fl Oz (Pack of 12)
Great variety of flavors, doesn’t make you jittery from caffeine, perfect size and great taste
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Reviewed in the United States on May 30, 2026
★★★★★ 4
cleaner caffeine hit, some flavors better than others
Flavor Name: Variety Pack, Size: 12 Fl Oz (Pack of 12)
Been going through this variety pack for a few weeks now and it's become a solid pre-workout alternative. 180mg caffeine from natural sources hits noticeably cleaner than the usual energy drinks — no jittery spike and crash. The sparkling texture is satisfying without being overly carbonated.
Flavor-wise the variety pack is hit or miss. A couple of the flavors are genuinely good, a couple I'd skip if buying singles. The zero sugar formula doesn't have that artificial sweetener aftertaste that ruins a lot of "healthy" energy drinks, which I appreciate.
The price is a bit higher than standard energy drinks but you're paying for the cleaner ingredient list. If you care about what's in your drinks, it's worth it.
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Reviewed in the United States on April 13, 2026
★★★★★ 5
The best ever!
Flavor Name: Variety Pack, Size: 12 Fl Oz (Pack of 12)
I lost my taste for coffee so I ended up trying several different energy drinks. Most of them are very, very unhealthy. However, I found this one “Bloom“. It is refreshing and has enough caffeine to keep me happy. The flavor is great except that I wish they would not use sucralose. It is not necessary.
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Reviewed in the United States on May 12, 2026