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Description
Human CHUK ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing 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 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 Conserved Helix Loop Helix Ubiquitous Kinase (CHUK) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Conserved Helix Loop Helix Ubiquitous Kinase (CHUK) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Conserved Helix Loop Helix Ubiquitous Kinase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The conserved helix-loop-helix ubiquitous kinase (CHUK), also known as nuclear factor kappa-B kinase subunit alpha (IKK-α or IKK1), is a protein kinase encoded by the CHUK gene. It is part of the IκB kinase complex and plays a key role in regulating the NF-κB transcription factor. However, it has numerous other cellular targets and is thought to function independently of the NF-κB pathway to regulate epidermal differentiation. It is a regulator of keratinocyte terminal differentiation and proliferation and plays a role in skin cancer. Activation of three major hydrogen peroxide-dependent pathways—EGF, FOXO1, and IKK-α—occurs during wound-induced epidermal keratinocyte migration, adhesion, cytoprotection, and wound healing. It regulates keratinocyte migration by monitoring the redox environment following wounding. It is sulfenylated at a conserved cysteine residue in the kinase domain, which correlates with inhibition of EGF promoter activity and increased EGF expression, suggesting that it stimulates migration through dynamic interactions with the EGF promoter, depending on the intracellular redox state. It also regulates cyclin D1 in an NF-κB-independent manner. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.5 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
Exceeded expectations!
Size: Mini Size
This ball launcher has been the best thing we ever bought. We have a golden retriever who of course, like most goldens, is ball obsessed. I have thrown my fair share. I decided to see if my golden was smart enough to use this so we pulled the trigger. It took 3 days of on an off training and BAM! We are in business! This launcher has been a CHAMP! We've had it a month now and it has survived over and over of constant use. The balls are covered in pure slobber and nastiness but the machine keeps going. We do keep it plugged in and only use it indoors. Sometimes the ball does get stuck but it's easy to push it back through. The only complaint I have, and it's no fault to the company, but we are already on our 2nd pack of replacement balls. Honestly, it's been worth it. I probably should have purchased the bigger size but I didn't want to invest in it if he wouldn't use it. Definitely regretting it now!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 27, 2026
★★★★★ 5
Best system ever !!
My 12 yr. old Toy Poodle was already trained to a 'piddle pad' , so this system was easy to introduce. I simplly placed her piddle mat in the sturdy sided piddle box and she used it immediately. With a straight lower pad holder, she was peeing at the side and I was constantly having to clean the surrounding floor . Now, it is all contained and she is happy with her new 'piddle place'. And, I am very happy too !!
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Reviewed in the United States on June 3, 2025
★★★★★ 5
Great!
Took my dogs a couple days to get used to using it but works well, great for apartment living !
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Reviewed in the United States on February 28, 2026
★★★★★ 5
Perfect for dachshund
This is the absolute perfect size and with sides. I have one without sides and it's great but my dog has decided to stand on the edge and sometimes pee goes off or poop. Also he will be excited when I get home and try to pee fast and run and it gets on the floor. This will slow him down a little. The grid is almost too small and it holds pee. I have to wipe it everytime with a paper towel. I have a 10lb dachshund. He is long and he has plenty of room. I have 23x23 pads. I fold one side of the extra plastic down on the pad and it fits perfectly. I highly recommend this one!!!
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Reviewed in the United States on March 6, 2024
★★★★★ 1
Not for small dogs
Not meant for small dogs at all. holes are just the size for the nails to slip in and get stuck. my dog tried to walk off pad and ripped his nail off! I do not recommend
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Reviewed in the United States on November 13, 2025
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