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Description
Rat IFNa 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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. 5. 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. 6. 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 200 pg/mL). Then dilute to the following concentrations: 200 pg/mL, 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 200pg/mL standard working solution into the first EP tube and mix thoroughly to make a 100pg/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 Interferon Alpha (IFNa) capture antibody. 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 Interferon Alpha (IFNa) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Interferon Alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interferon alpha (INN), or HuIFN-α-Le, marketed as Multiferon, is a drug composed of natural interferon alpha (IFN-α) obtained from the white blood cell fraction of blood after induction with Sendai virus. Interferon alpha contains several naturally occurring IFN-α subtypes and is purified by affinity chromatography. It is a mixture of several proteins that all possess the typical structural, serological, and functional properties of natural interferon alpha (IFN-α). The major subtypes identified are IFN-α1, IFN-α2, IFN-α8, IFN-α10, IFN-α14, and IFN-α21. IFN-α2 and IFN-α14 are glycosylated. | |||||||||||||||||||||||||||||||||
| 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 | 3.12-200 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.7 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 5
MUST HAVE!!! Dog is obsessed!!!
Color: Aqua
100 PLUS STARS!!! We have a very VERY active pup (6 yrs old 🤦♀️🤦♀️)… he has destroyed EVERYTHING we have ever given him. So I ordered this AND the replacement shell. He has chewed the ball extensively but it is in PERFECT shape, no need for the replacement one YET.. I can not rave about this enough - he LOVES it, we love watching him with it and he won’t let it out of his site at all!! If we touch it he gets upset.. so when we charge it, we put the shell back together and give him that.. which reminds me - make sure you have that screwed TIGHT.. somehow he was able to get it apart.. but once we started tightening it he hasn’t been able to.. thinking about it?? GET IT!! You won’t be disappointed and neither will your dog!!
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Reviewed in the United States on November 15, 2025
★★★★★ 4
An expensive ball to think about
Color: Aqua
Went with the foam ball because I have hopes that my dogs teeth will sink in enough to be satisfying without having to shred it like a tennis ball. So far, so good. I chose this one because the of the outer shell replacement possibility; assurance that 30 dollars towards a ball would have an extended life option for half the price if needed.
It is slightly bigger than a tennis ball and big enough for my dog to be unable to get a strong clench and crush it. He does try though and it's holding up.
All of the options and modes are great.
However, I wish it acted more of a bait ball. Going through all three of the modes, I can't say any have sparked a wild chase.
It will bounce intermittently or once touched. The intervals are long enough to lose my dogs interests. Short intervals make the ball the easy loser every time.
Better on hardwood but pathetic on carpet.
It will roll but not far or very fast.
The only positives from this may be the battery conservation but when he stops playing with it, it is still rolling around the room. I think it takes 20-30 minutes of inactivity for sleep mode.
With this, the charge will last about a day of play.
It does charge quickly.
It has passed the tough chewer test with one who can chew through "tuff" toys in no time but I wish it was more of a challenge.
After telling my sister i purchased one, she said she would've given me hers because her dog had lost interest. Go with your hunch and find another game to invest in if you have trepidations of this being this case.
I won't deduct anymore stars because the product is as advertised. Only one deducted for the price.
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Reviewed in the United States on September 19, 2025
★★★★★ 5
So far so good
Color: Aqua
For the past 2 years I’ve been looking for an interactive ball our dog won’t break within days.
I appreciate that it’s all foam, without any ropes or plastic pieces he can chomp on.
I don’t let him use it as a chew toy but after catching it he does like to chew on it for a little bit, and it’s held up nicely after a month.
Would recommend
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Reviewed in the United States on January 7, 2026
★★★★★ 2
if vibration excites your dog then it's okay
Color: Aqua
They're okay bit just basically vibrate. Was hoping they would roll around and their own but just really vibrate so my dogs didn't get much from it.
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Reviewed in the United States on November 29, 2025
★★★★★ 4
Please make a smaller one!! LOVE 💕
Color: Aqua, Color: Aqua
Giving a 4 star only because ….you dont have a smaller one!!
My dog loves these!!!
PLEASE PLEASE PLEASE!!! Make a smaller one!!!….. Just like this one!!!!!!
Made very well!!
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Reviewed in the United States on January 4, 2026