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Description
Human CXC motif 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 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 Breast and Kidney Expressed Chemokine/Chemokine (CXC motif) ligand 14 (BRAK/CXCL14). 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 Breast and Kidney Expressed Chemokine/Chemokine (CXC motif) ligand 14 (BRAK/CXCL14) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Breast And Kidney Expressed Chemokine;Chemokine ligand 14 (BRAK;CXCL14) ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | BRAK/CXCL14 is a small cytokine belonging to the CXC chemokine family. Mature CXCL14 shares many conserved features of the CXC chemokine subfamily, but also exhibits some differences, such as a shorter N-terminus and five additional amino acids in the region between the third and fourth cysteines. It is expressed at high levels in many normal tissues, and its cellular origin is thought to be fibroblasts. This chemokine is chemotactic for monocytes and can activate these cells in the presence of the inflammatory mediator prostaglandin E2 (PGE2). It is also a potent chemoattractant and activator of dendritic cells, implicated in their homing, and can stimulate the migration of activated natural killer (NK) cells. It also inhibits angiogenesis, likely by blocking chemotaxis of endothelial cells. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.4 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 5
Love These Balls
Color: H) 2.5" 3-Pack (Beef), Color: H) 2.5" 3-Pack (Beef)
These are great balls. My Border Collie can destroy a tennis ball in a couple of hours and the Kong brand ball is heavier than I like to play with in the house. These balls hold up to her biting and chewing but are lite weight and softer for bouncing off the walls.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 7, 2026
★★★★★ 3
Seems durable - but the scent is not the best
Color: B) 3" 2-Pack (Bacon)
I don't know if I'd call the scent of this "bacon," more like rubber with a hint of bacon. It's very strong. I gave my dog one of the balls & put one in a cabinet where I keep his treats & a few other unrelated odds & ends. I now hold my breath when I open that cabinet b/c good lord, the smell. I'm tempted to just give that one to the dog too, just because it'll air out eventually & then everything else in that cabinet won't smell bad.
My dog feels so-so about it. He'll briefly chase after it & chew on it, but loses interest in it pretty fast. My rather hyperactive cat also plays with it from time to time, as he seems to have more interest in most dog toys than the dog these days, so there's that. (Then again, he thinks a wide variety of random items around the house are loads of fun to toss about, so it's possible his opinion is to be taken with a grain of salt. Between the brief periods of one of these balls being chewed on & clawed at, it's stayed completely intact. I think if you have a dog who simply likes to play fetch or chew on rubber balls in general, this one would stand up to the task. But if you're expecting it to be greeted with the enthusiasm as a bacon grease covered ball might be - I'd adjust your expectations.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 19, 2023
★★★★★ 5
My dogs favorite toy so far in two years
Color: C) 4" 2-Pack (Bacon)
I use this with my medium sized dog. These are large for him but that's what I want so I don't ever have to think about them being too small or a swallow hazard. He has been through A LOT OF TOYS in the last couple years, but nothing, NOTHING has he loved as much as these.
His favorite used to be squeaker balls. Then his favorite was crackle balls. But he will gladly ignore both for this one. I don't know if it's the bacon flavor or what, but he go nuts for it.
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Reviewed in the United States on May 5, 2026
★★★★★ 5
Bacon? IDK but good stuff either way
Color: C) 4" 2-Pack (Bacon)
What is there to say? they're giant 4" rubber balls from the Chew King brand which is very durable brand. The best part is they don't roll under couches, side boards, etc cause they're just big enough.
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Reviewed in the United States on April 24, 2026
★★★★★ 5
Durable, bright and bouncy
Color: H) 2.5" 3-Pack (Beef)
Love these. Color is bright so I can find it in grass and dirt. Quality is good! Works well in a Chuck it. Love these holes for whistle sound as it’s in the air. No scent cost is awesome for all 3 in pack
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 30, 2025
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