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Description
Rat N-MID-OT 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 the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 4000pg/mL). Then dilute to the following concentrations: 4000pg/mL, 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 4000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 2000pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 utilizes 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 N-terminal mid-segment osteocalcin (N-MID-OT). 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 N-terminal mid-segment osteocalcin (N-MID-OT) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat N-terminal mid-segment osteocalcin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Osteocalcin, also known as bone R-hydroxyglutamic acid protein (GLa protein), is a 49-amino acid protein. It is a non-collagenous acidic glycoprotein and a vitamin K-dependent calcium-binding protein. It is primarily synthesized by osteoblasts and odontoblasts, with some synthesis by proliferating chondrocytes. In its uncarboxylated form, osteocalcin acts as a hormone in the body, sending signals to the pancreas, fat, muscle, testicles, and brain, playing a key role in regulating bone calcium metabolism. | |||||||||||||||||||||||||||||||||
| 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 | 62.5-4000pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 1072 reviews
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Product Reviews
★★★★★ 5
Upsize
Color: Pink, Size: XX-Large
Knowing to upsize before I bought the shirt was a good thing to know. Otherwise, it is a very nice shirt
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 30, 2026
★★★★★ 5
As I expected, Manly
Color: Blue, Size: Medium
this is a beautiful well made shirt
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 8, 2026
★★★★★ 5
No flaws, not high end
Color: Pink, Size: Medium
I nabbed this for my in shape 14 year old son who had an upcoming wrestling banquet. It fit very true to size for him however as a low-mid grade dress shirt it had little flex or mobility across the chest and that’s ok, this is an entry level dress shirt and perfect for this and similar events.
The color was vibrant and well saturated, and the fabric itself was thin enough for some transparency but thick enough to be considered medium weight. It would not be a cooling dress shirt.
It did not have a soft hand, rather a sturdy rougher feel akin to a dress shirt a restaurant server would wear and laundry frequently which is also ok because it is exactly what a polo in this range should be. If you are looking for a flexible chest and soft touch you will need a better fabric blend and you pay a lot more for that.
The buttons were nice and collar looked good.
I would recommend this for any average event or occasion or even for work. If you are looking to impress on an interview or a date or during high heat you might need to look elsewhere.
For what it is it’s perfect. Buyer and shirt need to remember this and keep it in its shirt lane.
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Reviewed in the United States on April 18, 2026
★★★★★ 4
Good shirt, not great, but it was never trying to be
Color: Navy Blue, Size: Medium
Let’s start with the fabric. The 50/50 cotton-polyester blend sounds like a compromise, and I suppose it is — but it’s a good one. You get enough cotton to feel like an actual shirt and enough polyester to keep things from turning into a sweat tent in the heat. It holds its shape after washing. No complaints.
The collar is a genuine highlight. The pin tips keep the points flat and symmetrical, which is a small thing until you’ve owned a shirt where the collar curls after two washes. You’ll notice it. There’s also a hidden plaid print under the collar — a subtle detail that’s only visible when unbuttoned. Unnecessary, but charming.
Fit is true to size. Order what you normally order.
It’s versatile enough to tuck in for the office or wear loose on the weekend. Not going to replace your best Oxford. But at this price, it was never trying to.
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Reviewed in the United States on March 27, 2026
★★★★★ 5
No bleed through of t-shirt color beneath. Very hard to achieve, but Alimens does it.
Size: X-Large, Color: All White
At the price point of this shirt, i'm super impressed with the overall quality, fit and appearance. I have had short sleeve white from others in the past and what sets this apart from them is this ... this needs no t-shirt underneath. if you did choose to add a t-shirt, the t-shirt color will not bleed through the white optics. I am daily shirt and tie guy, who loves button downs. I own many VH brand shirts and this Oxford is sized exactly as a VH. Will be ordering more!!
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Reviewed in the United States on May 14, 2026
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