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Description
Human CEBPb 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 and centrifuge 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. Cell Supernatant: 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., 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 the 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. Preparation before testing: 1. Please remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Preparation of gradient standard working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, and then gently mix (concentration is 10ng/mL). Then dilute to the following concentrations: 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.3125ng/mL, 0.15625ng/mL, and 0ng/mL. Serial dilution method: Take 7 EP tubes and add 500uL of universal diluent to each tube. 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 step to mix thoroughly. 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 a capture antibody against CCAAT/Enhancer Binding Protein Beta (CEBPb). 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 CCAAT/Enhancer Binding Protein Beta (CEBPb) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human CCAAT/Enhancer Binding Protein Beta ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | CCAAT enhancer binding protein beta, also known as CEBPb, is a protein encoded by the CEBPb gene. The protein encoded by this intronless gene is a bZIP transcription factor that can bind to certain DNA regulatory regions as a homodimer. It can also form heterodimers with the related proteins CEBP-alpha, CEBP-delta, and CEBP-gamma. The encoded protein is important in regulating genes involved in immune and inflammatory responses and has been shown to bind to the IL-1 response element in the IL-6 gene and to the regulatory regions of several acute phase and cytokine genes. In addition, the encoded protein can bind to promoters and upstream elements and stimulate the expression of type I collagen genes. It is essential for the normal function of macrophages, an important immune cell subtype. People who cannot express it have macrophages that cannot differentiate (specialize) and are therefore unable to perform all of their biological functions - including macrophage-mediated muscle repair. [8] Observational work has shown that its expression in blood leukocytes is positively correlated with human muscle strength, emphasizing the importance of the immune system, especially macrophages, in maintaining muscle function. | |||||||||||||||||||||||||||||||||
| 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, cell supernatant, tissue homogenate, etc. |
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4.2 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 5
Durable
Color: S6-muti-color
The balls are great for chewers. The squeaky didn’t last long but they are perfect for the ball launcher.
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Reviewed in the United States on May 18, 2026
★★★★★ 4
The squeeker isn't durable, but the balls themselves are.
Color: S6-muti-color, Color: S6-muti-color
I have two 70+lb, ball obsessed dogs (a field bred Golden Retriever, and an Old Time Scotch Collie), and these balls are living up to the abuse they're receiving.
In the fall we always lose a ball or two in the yard due to getting lost in leaves or longer grass, so I always try and buy a multi pack at the start of the season, so went with these this year. I'm glad I did. Great price for a 6 pack, they fit the medium Chuck-It launcher (a little loosely, but they throw just fine), have great bounce, they FLOAT (which means I didn't lose any at the beach), and they're super durable to the dreaded "mouth squish." The squeaker dies pretty quick, like, within a day, but it isn't the kind that can fall out or choke your dog, so no worries there.
The boys are happy, I'm happy, my wallet is happy - I've already ordered another pack.
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Reviewed in the United States on November 2, 2024
★★★★★ 5
Good balls
Color: S6-glow
My dog is a chewer, and these have held up for close to a year actually a pretty good product
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Reviewed in the United States on May 27, 2026
★★★★★ 3
Disappointed
Color: S6-muti-color
These were great for about 20 minutes then they stopped squeaking...we have 2 left that I have saved but the other 4 do not squeak anymore...disappointed as my dog loves to squeak things...would not buy these again...My dog is only an 8lb dog too...but he can still play with the balls without the squeak...
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Reviewed in the United States on April 26, 2026
★★★★★ 5
Great Value for a ball my dog loves!
Color: S6-muti-color
I wish these ball were in the subscription program! My dog gets sooo excited by the new ball, but her interest in things lasts for a few days. She kills the squeak within 11minutes, but the squeaker stays in place rather than becoming a chocking or swallow hazard, and she still loves to chew it. Next, we must throw the ball approximately 723 times the first day, possibly 496 times on day 2, and just 3- 4 on day 3, at which point she will chase the ball before hollering "it's over here if you need it," while she checks on the chipmunk den. We currently have 17 of these balls in our yard (because we have given several leftovers to a less discriminating doodle next door). These balls hold up really well, get her extremely active for the first couple of days, and are a much cheaper than doggie daycare (after an active morning with a new ball, she is happy to chew on it and sleep much of the day). I always have these on hand!
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Reviewed in the United States on November 22, 2025