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Description
Human 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: 1. 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: 1. 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. 2. 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. 3. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. 4. 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 Enolase 1 (ENO1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Enolase 1 (ENO1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Enolase 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | α-Enolase (ENO1), also known as enolase 1, is a glycoenzyme 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 its monomeric form, α-enolase functions as a structural crystallin (tau-crystallin). Alternative splicing of this gene results in a shorter isoform that has been shown to bind to the c-myc promoter and function as a tumor suppressor. α-Enolase has also been identified as an autoantigen in Hashimoto's encephalopathy. ENO1 is one of three enolase isoforms, the others being ENO2 (ENO-γ) and ENO3 (ENO-β). Each isoform is a protein subunit that can hetero- or homo-aggregate to form αα, αβ, αγ, ββ, and γγ dimers. The ENO1 gene spans 18 kb, lacks a TATA box, and possesses multiple transcription start sites. A hypoxia-responsive element (HFE) is found in the ENO1 promoter, enabling the enzyme to function in aerobic glycolysis and contribute to the Warburg effect in tumor cells. As an enolase, ENO1 is a glycolytic enzyme that catalyzes the conversion of 2-phosphoglycerate to phosphoenolpyruvate. Within cells, ENO1 is primarily localized to the cytoplasm, although an alternatively translated form is localized to the nucleus. The nuclear form, also known as MBP1, acts solely as a tumor suppressor by binding to and repressing the c-myc oncogene promoter; it lacks the glycolytic activity of the cytoplasmic form. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.6 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 3
Nice Edger as long as the blade doesn't come loose -- and it will come loose, repeatedly!
Probably could've/should've rated this 2 stars yet it works well when it works. I purchased this edger to edge my drive and sidewalks and not for trenching. The edger arrived very quickly after I ordered it (I am not a Prime member so this was a nice surprise). Assembly was easy and straight-forward. The first use went smoothly and the edger performed flawlessly and provided clean, smooth edging. About 20 minutes into the second use I noticed that the edger was cutting and quickly determined that the blade was spinning freely -- fortunately the hex nut was still attached to the threaded shaft. I reviewed the manual that came with the edger, grabbed the correct sized socket and torque wrench. I removed the hex nut, the first washer, the blade and then the second washer. To install I followed the directions ensuring that the flats of the washer aligned with the flat on the shaft and tightened the hex nut to the specified 140 in-lbs. The edger worked well for about 10 minutes and the blade once again became loose. This time I replaced the original blade (which did not need to be replaced from a wear perspective) with a new B&D blade and new washers. Once again I followed the instructions and this time the edger performed flawlessly until about 5 minutes remaining in my edging when once again the blade came loose. my experience has most certainly been frustrating to say the least. one should not need to carry a socket and torque wrench with them and repeatedly need to tighten the blade. So, when the edger performs as intended it does an excellent job at edging; however I have no confidence that this edger will be able to perform as intended throughout repeated uses and more importantly has shown that it cannot complete my edging without requiring tightening the blade (which requires disassembly down to the second keyed washer). Potential buyers should be prepared for this experience and make your decision accordingly. Note: I tried contacting black and decker via twitter (@blackdeckerhome) and never received a response; I could not find a separate customer service/support twitter account. Trying to contact customer service via the phone these days is an exercise in patience to say the least.
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Reviewed in the United States on November 11, 2022
★★★★★ 5
Trimmer results
Was alittle hesitant with this being B&D
But I was surprised how great it really is.
By far way above my expectations. Works like a charm and does a great job. Chose electric because I really don't have other than a driveway to do. But it really did good around the flower beds.
Cord limits you alittle but nothing to cry about. Quiet also but still use ear plugs.
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Reviewed in the United States on May 9, 2026
★★★★★ 5
Light weight and very functional
Incredibly light and functional, this trimmer forks with either string or resin blades. I personally prefer the blades. At the time I bought the unit I couldn’t locate replacement resin blades. I finally called Black and Decker and they sent me two packages of the replacement blades for free! The shield keeps trimming from being a messy horror show. No more back soreness because of the ligh weight of the unit. I’m very pleased with both the product and Black and Decker.
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Reviewed in the United States on July 22, 2025
★★★★★ 5
Espectacular
Es genial, tiene una potencia grandiosa.
Me encantó, la recomiendo 100%
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Reviewed in the United States on April 22, 2026
★★★★★ 4
Needs improvement
This trimmer really looks nice and it comes with some great attachments but what I found is that if you're going to switch from the line attachment to the little plastic attachment you have to sit somewhere where you can lay the item down because you have to unscrew one to put on the other. There is no way to easily switch from line trimming to attachment trimming without using the device that unhooks the screw. In my opinion that is really a detriment to the beauty of the overall working of this item the weight of it is good, it cuts good it automatically feeds the line out the color is nice and the price is great but then you have the issue of changing the line back and forth if this was improved I think I would definitely give it five stars I'm sorry but improvements need to be made.
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Reviewed in the United States on August 14, 2025
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