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Description
Human CLEC4G 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 the collected cells three times with ice-cold PBS and resuspend them 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 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 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 c-type lectin domain family 4 member G (CLEC4G) capture antibody. 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 c-type lectin domain family 4 member G (CLEC4G) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human c-type lectin domain family 4 member G ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | C-type lectin domain family 4 member G (CLEC4G), also known as LSECtin (liver and lymph node sinusoidal endothelial cell C-type lectin), is a member of the second subgroup of the C-type (Ca2+-dependent) lectin superfamily. It is an approximately 40 kDa, single-pass, type II transmembrane glycoprotein with a length of 293 amino acids (aa). It contains a short N-terminal cytoplasmic tail (aa 1-31) and a 21-aa transmembrane region. Its extracellular region includes two N-linked glycosylation sites (a73 and 159), a coiled-coil neck domain (a96-136), a C-type lectin-like domain (CTLD) found in DC-SIGN and DC-SIGN receptors (a165-289), and a C-terminal Ca2+-dependent carbohydrate recognition domain (C-type CRD). | |||||||||||||||||||||||||||||||||
| 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 homogenates, cell lysates and other biological fluids |
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4.0 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 3
Almost great! But falls a little short
Size: Large, Color: Wine Red
It’s a great looking pea coat, but the stuffing/insulating material appeared to be lacking. Granted price is nice so one can only expect so much. I was willing to overlook this and then I tried it on - the fit was great except for the sleeves. I wear 32-33 sleeves and these must be 35-36. They swallow up my hands. It’s bummer because the maroon/wine color is so nice. I’ll be returning unfortunately
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 10, 2024
★★★★★ 5
Great quality :-)
Size: X-Large, Color: Black, Size: X-Large, Color: Black
So I purchased a Nother pea coat from a box store that didn’t quite fit right and was not very flattering, so I returned it.
I’m 6 foot tall 210 pounds with very wide shoulders and long arms so I decided to get an extra large.
It’s slightly loose around the waist section, but I think that’s supposed to be that way considering you may wanna wear layers underneath.
I may possibly get it tailored a little bit tired her, but the chest and arms fit great and the length is perfect.
Great quality for the price and definitely a nice piece to the collection!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 1, 2023
★★★★★ 5
Great jacket!
Size: Medium, Color: Green
This jacket is super stylish, fits perfect, and is extremely warm. I gave this as a gift and the person loved it. It seems like a great value and looks like it will last a long time!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 25, 2025
★★★★★ 5
Excellent quality - great buy! Looks sharp!
Size: Large, Color: Gray, Size: Large, Color: Gray
Great quality - this is a medium weight coat. Perfect as a car coat but too thin for cold winter days if you had to be outside for long periods of time. The fit is just perfect. Purchased a size large for a man 5’6 175 lbs with a small belly so peacoat double breasted look - looks great! Lining is standard and the grey color is very appealing. This is soft feeling coat made to look like wool without the heavy rougher wool feel. Only issue is that the coat comes packaged in a small bag and so when you open it the coat it totally wrinkled. You can knock out the wrinkles in the dryer with a wrinkle release dryer sheet or use a garment steamer but honestly the dryer trick is the key.
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Reviewed in the United States on February 12, 2025
★★★★★ 4
Long enough sleeves for a tall guy
Size: Large, Color: Green
I’m 6’3” and skinny, and was very impressed to find the sleeves were long enough for me in a large! The overall fit of the coat is boxy or straight, while I would have preferred a slimmer cut, I’m just so happy that the sleeves fit!
The liner is nice, interior breast pockets on both sides are super convenient, and I love the green.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 15, 2026
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