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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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4.2 ★★★★★
Based on 6 reviews
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Product Reviews
★★★★★ 4
Slow, sorry but good
Format: Kindle
This was a slow moving book. Lots of character pov chapters, lots of superfluous descriptions and endless courtly appearances stalled this book to start. Once you get into the heart of the story, it takes off. Before you know it the book is done. My favorite character is Reyna. She is so strong. She is true to herself. She gets into a lot of trouble with her headstrong ways but it's entertaining. I have high hopes for Lorcan. He is honorable to a fault. Thane had turned out to be better than I thought but i still don't like him. Eislin is useless. Great plot twists at the end. I'm looking forward too book 2.
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Reviewed in the United States on June 6, 2020
★★★★★ 5
Very detailed! Beautiful world building! Strong Heroine!
Format: Kindle
Starts off a little slow and confusing with different POV’s, but starts to all come together towards the middle to make an elaborate plot line and makes it all worth it. Beautiful world building and attention to detail as well as great writing. The cliffhanger was gut wrenching! Can’t wait for the next book!
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Reviewed in the United States on March 4, 2020
★★★★★ 4
Interesting
Format: Kindle
This was a very captivating book once you got into it thoroughly. But the third person perspective was a bit hard to get used to. But as you got into it and followed the different characters, it was interesting and filled with intrigue, conflict and forbidden love.
I can’t wait to read the next one and to complete the series.
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Reviewed in the United States on November 12, 2022
★★★★★ 5
OBSESSED with this start to the series + incredible narration!
Format: Audiobook
5⭐️ 3🌶️ 5🎧
Publisher: Podium
Narrators: June Deborahae, Vincent Skye, Ace Bentley
Narration type: multicast, dual style
Writing perspective: first person, multi POV
Standard length: 9:28
Listening speed: 1.9x
Tropes: why choose, sports, new adult/college, forced proximity, found family, one bed, disability, MH + neurodivergent rep
📖 I have always been a why choose lover and this is def going to be a new fav series, I can tell already! I absolutely LOVE Abby’s writing! Joey is such a great FMC — she is strong, but has a past that has pain, making her a complex and layered character with some mystery. The guys are amazing as well — they are all different and react in such varying ways to Joey showing up on the scene! If you are going in expecting a full on dark romance with kidnapping vibes, this is not it. This is definitely a romance with darker themes, but these guys treat their girl well, though they may be misguided at times (cough, Decker, cough 🤣). One of my absolute fav things about this book is the incredible representation — two of the guys deal with chronic pain (RA + lupus), another is on the spectrum, and Joey has anxiety that can lead to panic attacks and more severe issues. As someone who deals with chronic pain (an autoimmune arthritis in the RA family) I love to see characters like me in books! I have no doubt by the end of the series I am going to love all the guys equally, but right now I am falling HARD for Kylian (who doesn’t love stats daddy?!) and Locke is firmly in second place. Decker and Kendrick have some work to do, but I’m sure this is all according to Abby’s master plan. And wowwww that cliffy was a painful one! I’ll be starting book 2 in T-minus 3 seconds!
🎧 This audio was phenomenal! All three narrators did an incredible job — the casting was absolutely on point!! June deBorahae narrated the majority of the book as most is from Joey’s POV and she was magnificent. Her transition from narrator internal voice to dialogue was always very clear, as were her transitions in dialogue between all characters which was an impressive feat with the number of characters she had going on. She truly blew me away as Joey though. The emotion she showed was incredible — going from tough and ready to go head to head with anyone, to exceedingly compassionate and kind, to anxious and sucked into memories of the past.
Vincent Skye voiced Decker, Locke, and Kylian and did a great job with all of them. Each was voiced in a way that was fitting for their character and personality. I especially love the way he portrayed Kylian.
And last but not least, Ace Bentley narrated the part of Kendrick to perfection. His portrayal was so fitting for his tough and grumpy ways. While Kendrick only had one POV chapter in this book, Abby tells me we can expect more from Ace as Kendrick in the next one!!
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Reviewed in the United States on March 19, 2024
★★★★★ 4
So good!
Format: Kindle
This was a great dark romance. There was lots of suspense and good spice. 🌶️🌶️🌶️. It ended on a cliffhanger so I’m looking forward to reading the next book.
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Reviewed in the United States on January 31, 2026