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Description
Human IGKV1D-12 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. 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 utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with an immunoglobulin kappa variant 1D-12 (IGKV1D-12) 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 immunoglobulin kappa variant 1D-12 (IGKV1D-12) in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Immunoglobulin kappa variant 1D-12 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Immunoglobulin kappa variant 1D-12 (IGKV1D-12) is a protein-coding gene. An important analog of this gene is IGKV1-12. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 5
It's hot.
Color: Silver
It's hot. I like it.
I make my tea in a 20 oz mug.
Invariably it gets cold and I need to reheat it. Someone suggested I order a mug warmer. I chose this one.
It has 3 heat settings and a 12 hr timer with auto turn off. What more could I ask for?
Except that it has a low profile and easy to read and set settings, it does.
Prefect.
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Reviewed in the United States on May 19, 2026
★★★★★ 5
Effective and easy to use.
Color: Silver
Keeps my coffee and tea hot while I work at my desk. Easy to set temperatures, and to change them for different beverages.
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Reviewed in the United States on May 17, 2026
★★★★★ 5
The perfect coffee mug warmer!
Color: Wood
If you need a coffee cup warmer for your desk or work-station, this is it! I've been using almost daily for several weeks now and zero complaints thus far.
As with many-a-reviewer, the problem I was trying to solve is my coffee going cold long before I was done with it, leading to repeated microwave rewarm sessions. After reading through numerous reviews, articles on the matter, and forum postings, I settled on this product. My requirements:
- I did not want to use a specialized cup
- I did not want to have to find "flat-bottom" cups, I wanted to use any cup in my cupboard
- I wanted the coffee to stay hot, not just warm
- I needed auto-off, since I surely would forget to turn it off
- Idiot-proof controls
This product's description seemed to check all the boxes, and it delivered! I've been using for several weeks and it's been perfect. It works with my regular ceramic coffee mugs, and on high setting, keeps my coffee pretty hot (130F). All my ceramic mugs are concave bottom - look at picture in my review - so that's why it only gets that hot on High. A flat-bottom cup may only need Medium setting since it would have more contact area for heat transfer.
The faux-wood trim makes it look sorta classy, and although it is large, it doesn't look absurd on my desk, and I'd rather it be bigger than my cup, rather than too small. And I imagine I could use it to keep soup or other food warm on my desk, since I also take forever to finish food when I do eat at my desk. The faux-wood trim does not get hot-to-touch, so it also prevents burns from accidental touch. The surface is, of course, very hot but no little kids or cats in our house, so that wasn't a concern for me.
I'm glad I went with this 36W option, even though some articles I read indicated this was overkill. As mentioned above, I imagine I need the higher wattage heating element because I use regular concave-bottom coffee mugs. I have it plugged into surge protector that powers my tower PC and 4 UHD 27" monitors, and I haven't tripped anything yet. 36W is even less than what a fast-charging laptop charger would pull, so this doesn't seem excessive for a workstation.
FOOD SAFETY WARNING: As a former research lab assistant that grew bacterial cultures as one of my duties, I'd like to remind people to keep in mind that the danger zone for bacterial growth is ~40F to 140F, especially with dairy creamers. To test temperatures, I used a ceramic concave-bottom mug (starbucks mug in picture) and a metallic flat-bottom mug (green mug in picture). Here are the liquid temperatures I measured for this warmer:
- Ceramic convex-bottom mug, Warmer on High: 130F
- Metallic flat-bottom mug, Warmer on High: 175F
- Metallic flat-bottom mug, Warmer on Med: 150F
As you can see, the concave-bottom makes a significant difference in heat transfer, and thus the temperature of the liquid. Use common sense on how long you leave your coffee on the warmer. Coffee at 130F is probably fine for a couple hours, but if you're gonna be leaving it all day, you'll want it to be at 140F+, so use a flat-bottom cup. The metallic one I have got real hot to the touch, including handle and lip of cup, even at Medium, so you'll probably want to use a ceramic one.
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Reviewed in the United States on April 7, 2024
★★★★★ 5
My favorite desk item
Color: Wood
I love my coffee but it must be hot-every sip! I'd get up at work so often to go reheat my coffee in the nuker & don'tlike the microwave AND a hater coworker even commented to me that she noticed "how much I get up from my desk" so I thought, they gotta be making coffee hot plates for desktops these days & boom, they do!! I like the wood finish, right my aesthetic alley. The plate has 3 settings; high, medium & low. Even has an auto shutoff after..idk how many hours so built in safety. I've even put my ceramic cup of soup on there to keep it warm while I work through lunch a couple times & it worked for my soup on the high setting. Coffee goes to medium when I first get in since it cools on the car ride over & then down to low once it's hot again. Will be getting one for the home office here soon.
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Reviewed in the United States on May 2, 2026
★★★★★ 5
Mug warmer
Color: Wood
This is a well made mug warmer, heats up quickly and convenient to use. The heat ratio is high making it easy to select the level of heat one wants to have to enjoy their coffee, tea, hot chocolate, etc. The only default I have with this product is how to turn on the warmer. It's convenient but if something is sitting next to the warmer, the warmer turns on too easily. I would recommend the manufacturer use some type of switch to turn on, adjust and turn off the warmer due to the chances of heating something up that was not intentional!
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Reviewed in the United States on April 26, 2026