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Description
Mouse CXCL5 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. 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. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 4000pg/mL). Then dilute to the following concentrations: 4000pg/mL, 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 4000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 2000pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 CXC motif chemokine 5 (CXCL5) 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 CXC motif chemokine 5 (CXCL5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse C-X-C motif chemokine 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | CXC-motif chemokine 5 (CXCL5 or ENA78) is a protein encoded by the CXCL5 gene. It is a small cytokine belonging to the CXC chemokine family and is also known as epithelial-derived neutrophil-activating peptide 78 (ENA-78). It is produced in response to stimulation by the inflammatory cytokines interleukin-1 or tumor necrosis factor-α. It exerts these effects by interacting with the cell surface chemokine receptor CXCR2. | |||||||||||||||||||||||||||||||||
| 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 | 62.5-4000pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.5 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 4
DONT BUY THE ELEPHANT, ITS NO GOOD. but GR8 BRAND!
Color: Checkers - Elephant (Gray), Size: Large
The elephant toy had no trunk by the FIRST day! The elephant now looks like an angry Koala bear OR an angry trunk less elephant(whichever u want to say) I LOVE the donkey and dinosaur toys so I bought the elephant confidently, thinking I had nothing to worry about. I wouldn't have spent money on this knowing the trunk would be chewed off in a matter of minutes. My Bella usually chews but she has had her donkey and dinosaurs with the chew guard for MONTHS and there are NO TEARS OR RIPS even with her playing and chewing on them ROUGHLY all day. If you buy any of these type toys I DO highly recommend the donkey or dinosaurs ONLY. The elephant being chewed up in a matter of minutes really makes me angry that I ever spent the money on it. HAD I known it wouldn't be the high quality dog toy I'm used to from GoDog, I would've just bought a second donkey or dino! Now Bella doesn't get her new toy for the month (I try 2 buy her a new stuffed toy or 2 about 1 time a month whenever I have the extra money for it) I guess she will have to go without this month because I spent it on this now trunkless elephant/angry koala:( Whichever name you call it, it is now going to be trash.
UPDATE!
If you contact the GODOG company they will send one replacement toy for your pet! They also asked me my pet's size, breed and age in order to send the best possible toy! They sent Bella a snake that has held up Extremely WELL! It's great and so far is my favorite(and one of Bella's favorites) The snake has less sqeaky things in it so she doesn't have the drsire to tear it apart immediately to retrieve them by rippong it to shreds:) My updated rating is for the MANUFACTURER'S GREAT CUSTOMER SERVICE. The fact that they actually replaced it was good but the fact that they went that extra mile by researching the best toy match for my Bella was above and beyond! If you have a Godog toy that went bust right away, contact the manufacturer for a replacement. If you give them your dog's age, weight and breed they will even try to match your pets replacement toy with something that is better suited to your dog's chewing style. (Bella's style is destructive haha)
Good luck, ya'll!
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Reviewed in the United States on October 26, 2016
★★★★★ 5
My Dobermans favorite toy to destroy, and one of our recommended picks.
Color: Checkers - Elephant (Gray), Size: Large
Ok, here's an honest breakdown review. I have a 6 month old Doberman who shreds every stuffed animal and dog bed he's come across. He's one of the worst chewers I've owned. We have tried a WIDE variety of stuffed toys.
That said, this elephant is his favorite toy. I have ordered 4 of them so far, and just ordered another 4-pack on a buy three get one free deal. The elephant stands out above all the other stuffed creatures from goDog for me as to durability. Hence why I keep buying more. But they usually last about 2-3 weeks on average.
In our house, the poor creatures are subjected to vicious, crazy, shaking of the head, helicopter type rotation. It' so hard if it hits your bare arm or leg it actually stings especially when its wet. Then the old, put it between his legs laying down, tearing at the legs action. The limbs are usually the first things to go over time. But they are each almost self sealing, so you clean up as little fluff and keep using it. Eventually the pup reaches the main body and finishes it off.
For a normal light to normal chewer, one of two will get you a long way. For those of us resigned to having zoomie type manic chewers that need a constant supply of toys, 2-3 week durability is pretty sound too. I do not recommend the green cheaper monster. It lasted about 45 minutes. The free replacement took about 3 days. Hopefully this aids you in your decision!
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Reviewed in the United States on June 10, 2026
★★★★★ 5
Very durable for rowdy pups
Color: Checkers - Elephant (Gray), Size: Large
This toy has outlasted any other dog toy I have ever bought. I have one large and one medium dog who share toys, fight over toys, chew them to death and still play with them. It might be because it isn't one of their favorites. I don't know. All I know is that our house is littered with dog toys in different states of destruction and this elephant toy has very little damage. It has definitely been in the dog toy rotation.
I had this toy for 2 months without any major damage. The day I wrote the review the dogs tore the leg off! I still think it is a good toy. Some toys lose the stuffing or squeaker the first day.
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Reviewed in the United States on May 18, 2025
★★★★★ 5
Great toy for a smaller dog
Color: Checkers - Dinos Bruto (Purple), Size: Mini
This is a small, firm, sturdy toy designed for chewing. It isn't a soft toy and it does not have a squeaker. My dog is only 8 lbs, and this is a perfect size for her. I discovered these smaller toys through my groomer. Other manufacturers make this size, as well. The "hard chew" part indicates that this toy will not be chewed to death easily. The oldest one that we have is still in good condition. It's a baby duck (also available on Amazon) and it's over a year old. The dinosaur is colorful, easy to throw, and a really cute. Every night, my dog gives these small toys a bath, cleaning them up by licking each one before going to sleep. It's as if they are her babies. Since she never had a liter, my vet said that they are about the size that her babies would have been, so she may see them as her babies. She treats them well. Every now and then they go" swimming" in the washing machine and come out looking bright and new again.
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Reviewed in the United States on January 2, 2026
★★★★★ 4
Updated, still going !! didn't last two minutes.
Color: Checkers - Elephant (Gray), Size: Large, Color: Checkers - Elephant (Gray), Size: Large
I hated giving this a 1 star, because it was so cute when I unpacked it today. I just gave it to my dog and in less than two minutes he has a hole ripped in it.! he loves it so I don't wanna take it away from him, but I'm sure it's going to be gone in another few minutes. Why can't someone make a stuffed animal for a dog that lasts like they say so on their listing? I really should take it away from him and send it back, but I don't have the heart.
I had to come back and give this a better review. it deserves at least four stars because even though he ripped holes in it, this one has lasted way longer than any other toy. He's actually still playing with it, and it still has some crinkle left to it so he didn't destroy the whole thing as quickly as he did other stuffed animals. I will certainly buy another one because at least it lasted a week so far!
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Reviewed in the United States on January 6, 2026