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Description
Rat ENO1 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 an alpha-enolase (ENO1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of alpha-enolase (ENO1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Alpha-enolase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Non-neuronal enolase (NNE), also known as enolase 1 (ENO1) or α-enolase, is a glycolytic enzyme expressed in most tissues and one of the enolase isozymes. Each isozyme is a homodimer composed of two α, two γ, or two β subunits and functions as a glycolytic enzyme. In addition, α-enolase, as a monomer, functions as a structural crystallin. Alternative splicing of this gene results in a shorter isoform that has been shown to bind to the c-Myc promoter and act as a tumor suppressor. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.2 ★★★★★
Based on 108 reviews
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Product Reviews
★★★★★ 5
Great Little Grinder — Solid Build & Easy to Use!
I’ve really enjoyed using this manual coffee grinder! For the price, it’s an excellent find. The adjustable settings work surprisingly well — I’ve been able to get a consistent grind whether I’m brewing for espresso or French press, which isn’t always the case with more affordable manual grinders. The ceramic burr gives a nice, even grind without overheating the beans, which helps preserve the flavor.
It’s easy to use and straightforward to adjust. The grind size clicks into place firmly, so you don’t feel like it’s going to slip or change while you’re using it. One fill gives me enough coffee for about two cups, which is perfect for my morning routine or when I’m brewing a small batch.
The build quality feels sturdy and durable, with a sleek look that doesn’t feel cheap. The handle turns smoothly and doesn’t wobble, which makes the whole experience feel solid and satisfying. I’ve taken it on a short weekend trip already, and it’s lightweight and compact enough for travel or camping.
If you're someone who loves the ritual of grinding your own beans or just wants a reliable backup grinder for when the power’s out, I’d highly recommend this one. Great value for a tool that feels like it’ll last!
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Reviewed in the United States on June 10, 2025
★★★★★ 5
Love it!
Perfect little manual grinder for my camp coffee! No power needed. Great value. Seems to be durable. Has different grinding settings. Easy to use. Definitely recommend!
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Reviewed in the United States on April 27, 2026
★★★★★ 3
Seems like a decent product, but it's not for me
First off, a small caveat: this is the first manual grinder I have used. I am trying to highlight things I think might be unique to this model, but some of these might be "that's just how manual grinders work" problems.
First and foremost, I can only cram about 20g of coffee into here at one time. If I'm making an 8oz cup, that tends to be enough -- but no larger than that. I also find it hard to load; last time I tried to just pour beans in most of them ended up on the floor. One scoop at a time is annoying if you are adding 20 grams of coffee, especially if you use a small scoop.
Also, I personally dislike that the body that holds the unground beans is opaque. I have to turn the thing upside down to determine if I have more left in there to grind or not. This is likely the same for other models but I don't think it makes for an ideal user experience.
Now, on the upside, it's compact, easy to use, and easy to clean. I don't know how to gauge the accuracy or consistency of the grind, but it seems to produce a somewhat finer grind than my electric burr grinder at what I would expect to be the same setting (20/40 on here is more fine than 14/28 on the other grinder, even though I would expect both of those to be "medium"). I'm someone who prefers slightly coarser grind, even for pour over, so this is another strike against it in my book.
I have only used it two or three times due to usually making larger cups that this can accommodate, and also not enjoying spending 5 minutes rotating my arms to get the coffee ground. So I also cannot speak to durability, but the plastic and glass both seem sturdy so I do expect it to last.
All in all, I love that it is lightweight, portable, and easy to clean, but I think I'm someone who prefers the ease of use that an electronic grinder can provide. And seriously, IDK how anyone is getting more than 20g of coffee into this thing at one time.
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Reviewed in the United States on January 6, 2026
★★★★★ 5
Perfect for Lattes
Size: 20 Ounces
Good for steaming 8-10 oz of milk. The nifty tool is good for scooping out your puck.
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Reviewed in the United States on February 19, 2026
★★★★★ 5
Almost blew up my microwave
Size: 20 Ounces, Size: 20 Ounces
TL;DR "A hilarious microwave mistake turned into a great home cafe upgrade. This 20oz cup is straightforward to handle, looks incredibly professional, and holds a level of heat that will literally burn you if you aren't careful."
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Bought this for homemade coffee brewing, just to spice things up. But as I go ahead to heat up my new shiny stainless steel milk frother cup in the microwave, whoopdy doo. Ya can’t put stainless in the microwave! Regardless, it’s still a straightforward frother cup to handle, making it a great dollar investment for upgrading my home cafe setup.
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1. Spotless Stainless Steel: Its sleek appearance made me feel professional in the hands of an amateur like moi. Great quality!
2. Temperature Retention: I accidentally burnt myself because it stayed so hot, but that is a major positive. Proving the insulation works perfectly and keeps the milk piping hot.
3. Size: The 20oz capacity is accurate, providing the perfect amount of space for frothing without any overflow.
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Reviewed in the United States on June 6, 2026