SKU: 22827410119

Mouse IL-13 ELISA Kit

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Description

Mouse IL-13 ELISA KitProduct Specification Usage Need to bring your own test equipment 1. Microplate reader (can measure the absorption value of 450nm detection wavelength and 540nm or 570nm correction wavelength) 2. High precision liquid dispenser and disposable suction head 3. Distilled water or deionized water 4. Washing bottle (spray bottle), multi channel plate washer or automatic plate washer 5. 500mL cylinder One, preparation before the experiment 1. Sample

Product Specification

Usage Need to bring your own test equipment
1. Microplate reader (can measure the absorption value of 450nm detection wavelength and 540nm or 570nm correction wavelength)
2. High precision liquid dispenser and disposable suction head
3. Distilled water or deionized water
4. Washing bottle (spray bottle), multi-channel plate washer or automatic plate washer
5. 500mL cylinder

One, preparation before the experiment
1. Sample collection and storage
① Cell culture supernatant: particles should be removed by centrifugation; Test the sample immediately. If the sample is not tested in time after collection, it is recommended that the sample be divided according to the dosage and stored in the refrigerator at -20 ° C to avoid repeated freezing and thawing. Samples may need to be diluted (1×) Dilute.
② Serum: Samples were collected using a serum separation tube (SST) and samples were left at room temperature for 30 minutes. The samples were centrifuged at 1000g for 15 min. Serum was immediately removed and tested immediately. If the sample is not tested in time after collection, it is recommended to repack according to a single dosage and freeze in ≤ -20℃ refrigerator to avoid repeated freezing and thawing. Samples may need to be diluted (1×) Dilute.
③ Plasma: Plasma was collected using EDTA, heparin, or citric acid as an anticoagulant, centrifuged at 1000g for 15 min within 30 min of collection, and tested immediately. If the samples are not detected in time after collection, it is recommended to separate the samples according to the single dosage and freeze them in &le. -20℃ refrigerator to avoid repeated freezing and thawing. Samples may need to be diluted (1×) Dilute.
2 Reagent preparation (Place all reagents and samples at room temperature for 15 minutes before use. It is recommended that all experimental samples and standards do double hole detection )
1× Preparation of washing solution: concentrated washing solution in the kit is 20× Mother liquor, diluted to 1&times with distilled water before use; Working liquid. Example: Take 10mL concentrated washing solution +190mL distilled water to 200mL, the actual operation can be calculated first, then make up.
②1× Dilution with buffer preparation: concentrate dilution in the kit with buffer 10× Mother liquor, diluted to 1&times with distilled water before use; Working solution. example: Take 3mL of concentrated dilution with buffer +27mL of distilled water to a constant volume of 30mL. In practical operation, the required dilution buffer can be calculated according to the dilution multiple of the sample, and then the preparation can be made.
③ Detection of antibody: the dry powder was centrifuged to the bottom of the tube, and 110uL dilution buffer (1×) was used. Dissolve and let stand at room temperature for 5 minutes to obtain 100× Mother liquor; Dilute to 1&times before use; Working solution. Calculate the desired volume by using 100uL per well. example: 10 Wells were used, then take 10uL of 100 times the working concentration of the test antibody, using dilution buffer (1×) Constant volume to 1mL, get 1mL of 1× The working concentration of the detected antibody.
④SA-HRP: SA-HRP is 40× Mother liquor, use dilution buffer before use (1×) Dilute to make 1× Working solution, 100uL required per well. example: used 10 holes, then take 25uL of 40× Mother liquor +975uL dilution buffer (1×) Constant volume to 1mL to obtain 1&times of 1mL; The working concentration of the detected antibody.
⑤ Chromogenic agent: according to 100uL per well, calculate the amount needed for this test, take out the corresponding volume of chromogenic agent, avoid light; The removed chromogenic agent is only used on the same day.
⑥ Standards: lyophilized standards with dilution buffer (1×) Redissolve, redissolve volume 1000uL, to obtain a concentration of 4000pg/mL standard mother liquor. Gently shake for at least 5 minutes, it is fully dissolved. Add 300uL dilution buffer (1×) to each dilution tube. . The standard mother liquor is diluted according to the picture below, and each tube must be fully mixed before pipetting to the next tube. The standard mother liquor without dilution can be used as the highest point of the standard curve (4000pg/mL), and the dilution buffer (1×) Can be used as the zero point of the standard curve (0pg/mL).

2, operation steps 
1. Ready for all the needed reagent and standard;
2 Remove the microplate from the sealed bag that has been balanced to room temperature, and put the unused slat back into the aluminum foil bag and re-seal it;
3. Add 300uL washing solution to the microplate, let it soak for 30 seconds, discard the washing solution and pat the microplate dry on absorbent paper, please use immediately do not let the microplate dry;
4. Add different concentrations of standard, experimental samples or quality control into the corresponding Wells, 100uL for each well. The Wells were sealed with plate adhesive and incubated at room temperature for 2 hours.
5 Suck the liquid out of the plate and wash the plate using a bottle washer, a multi-channel plate washer, or an automatic plate washer. Add 300uL of washing liquid to each well, and then suck the washing liquid out of the plate. Repeat 3 times. Every time you wash the plate, try to absorb the residual liquid to help you get a good test result. At the end of the last plate wash, please blot all the liquid in the plate or invert the plate and pat all the residual liquid in the absorbent paper;
6. Add 100uL detection antibody to each microwell. Seal the reaction Wells with sealer tape and incubate for 2 hours at room temperature;
7. Repeat the plate washing operation of step 5;
8. Add 100 ULSA-HRP to each microwell and incubate for 20 minutes at room temperature. Be careful to avoid light;
9. Repeat step 5 to wash the plate;
10. Add 100uL of color development solution to each microwell, incubate at room temperature for 5-30 minutes, pay attention to avoid light;
11. Add 50uL of termination solution to each microwell, and the color of the solution in the well will change from blue to yellow. If the color of the solution turns green or the color change is inconsistent, tap the microplate to mix the solution evenly;
12. Within 30 minutes after the termination solution is added, the absorbance value at 450nm is measured using a microplate reader and 540nm or 570nm is set as the correction wavelength. If the dual wavelength correction is not used, the accuracy of the results may be affected;
13 Calculation results: The corrected absorbance values (OD450-OD540/OD570), the compound reading were averaged for each standard and sample, and then the average zero standard OD value was subtracted. Standard curves were created by 4-parameter logic (4-PL) curve fitting using computer software. Alternatively, a curve can be generated by plotting the logarithm of the concentration of the standard against the logarithm of the corresponding OD value, and the best fit line can be determined by regression analysis. This process produces an adequate but less accurate fit to the data. If the sample is diluted, the concentration should be multiplied by the dilution.
Note: The standard curve data provided is for reference only. The sample content should be calculated according to the standard curve drawn in the same test.

3. Kit parameters
1. Recovery: Different levels of mouse IL-13 were incorporated into the cell culture medium samples, and the recovery rate was determined. The recovery ranged from 96 to 103%, with an average recovery of 99%.
2. Sensitivity: The minimum detectable dose (MDD) of IL-13 in mice was generally less than 0.089-0.740pg/mL. The lowest detectable value of IL-13 was calculated as the corresponding concentration based on the average of the zero absorbance values of 20 standard curves plus two standard deviations.
3. Correction: the ELISA kit by high purity recombinant e. coli expression correction by IL - 13 proteins in mice.
4. Linearity: Four different samples were mixed with high concentrations of mouse IL-13, followed by dilution (1×). Linearity was determined by dilutes the samples to the detection range.
Dilution ratio Mean value/expected value (%) Range (%)
1:2 102 100-104
1:4 105 102-108
1:8 107 103-111
1:16 107 100-113


5. Specificity: This ELISA can detect native and recombinant mouse IL-13 protein. The following factors in diluent (1 x) configured to the concentration of 50 ng/mL to detect cross reaction with IL - 13 in mice. To the interference of 50 ng/mL factor added to the middle scope IL - 13 reference substance in the restructuring of the mice, the interference of IL - 13 to test in mice. No significant cross-reactivity or interference was observed.

Recombinant human proteins Recombinant mouse proteins
IL-4 G-CSF IL-10R IL-3
IL-4 Rα GM-CSF LIF IL-4
IL-13 IFN-γ M-CSF IL-5
  IL-1α MIP-1α IL-7
  IL-1β TNF-α IL-9
  IL-2 MIP-1β IL-10

4, common problem resolution
1. The white board (no color), after the completion of color
NO. Cause Solution
1 Kit stored improperly; A mixture of different kit reagent Buy a new kit, pay attention to the storage conditions; Do not mix
2 Endow low temperature, short time If the temperature is too low, the incubation time is prolonged and the color development time is prolonged
3 Wrong addition or omission of reagents In strict accordance with the manual steps to add the correct reagents
4 Used to configure the solution container not clean, or there is something wrong with the water The use of clean containers and qualified distilled water
5 In the process of washing the plate, the soaking time is long, the number of washing the plate is too much, and the impact of washing the plate is large In strict accordance with the manual operation
6 The temperature of the reagent was not uniform All reagents were equilibrated at room temperature for 30 minutes
7 Detection of antibody and/or HRP concentrations was too low Do not dilute at will according to the instructions

2. Flower plate (blank, negative and positive controls were normal, but the OD value of sample Wells was significantly higher)
NO. Cause Solution
1 Fewer washing, inadequate Wash according to instructions
2 The substrate 3,3',5,5' -tetramethylbenzidine (TMB) is contaminated or exposed to metal ions or oxidants The use of clean containers and when making up qualified distilled water; Avoid light preservation
3 High incubation temperature and/or excessive incubation time Control incubation and the enzymatic reaction temperature and time
4 Sample did not change when the spear head, cause cross contamination Change the tip of each sample
5 Near the hole cross contamination Vertical clappers, using the appropriate legal pad, avoiding the hole into confetti
6 Samples are endogenous interfering substance Possible infectious agents were speculated and treated accordingly
7 Sample hemolysis, storage for too long, incomplete agglutination, contaminated by bacteria, blood vessels to add impact Avoid hemolysis, contamination, too long storage and other phenomena
Theory Double antibody sandwich enzyme-linked immunosorbent assay was used in this kit. Specific anti-mouse IL-13 antibody was precoated on a high-affinity microplate. After incubation, the IL-13 present in the sample combined with the solid-phase antibody and the detection antibody to form immune complexes. After washing to remove not combined with material, by adding horseradish peroxidase labeled chain mildew avidin (Streptavidin - HRP). After washing, the chromogenic substrate was added and the color was developed in the dark. Join terminated liquid termination reaction, in the 450 - nm wavelength (reference calibration wavelength of 540 nm or 570 nm) determination of absorbance values.
Composition
Component Size Once opened, diluted or heavy soluble reagent expiration date
Mouse IL-13 Microplate 1 Unused slats can be stored at 2-8 ° C for up to 30 days after being sealed back in aluminum foil bags with desiccants
Mouse IL-13 standard substance 2 Dissolves, calculate the dosage repackaging, can be in - 20 ° C storage for 14 days
Mouse IL-13 detect antibody 1 Enrichment dissolved after the volume, can be in 2-8 ° C storage for 14 days
40×SA-HRP 1 40× concentration can be stored at 2-8°C; 1 x working concentration is not recommended to save
10 x condense dilute with buffer 1 After opening, can be in 2-8 ° C stored for 30 days
Color-substrate solution 1
Stop buffer 1
20 x inspissation washing buffer 1
Sealing film 3 Room temperature storage, to avoid contamination, do not repeat use
Background Interleukin-13 (IL-13), a member of the chemokine family, plays a key role in the pathogenesis of allergy, cancer and tissue fibrosis. It is secreted by several helper T cell subsets, NK cells, mast cells, eosinophils, basophils and visceral smooth muscle cells. IL-13 inhibits the production of proinflammatory cytokines and other cytotoxic substances by macrophages, fibroblasts and endothelial cells. It promotes B cell activation, immunoglobulin class switching to IgE and upregulation of CD23/Fc epsilon RII. IL-13 signals through a receptor complex containing IL-13 R alpha1 and IL-4 R alpha. The compound is used as a type 2 receptor IL - 4. IL-13 ralpha2 binds IL-13 and prevents IL-13 from signaling through the IL-13R alpha1/IL-4 Ralpha complex. Il-13-bound IL-13 R alpha2 can directly promote tumor cell invasion and tissue fibrosis development.
General Notes 1. Please use the kit within the validity period.
2. Components of different kits and different batch kits should not be mixed.
3. If the sample value is greater than the highest value of the standard curve, the sample should be diluted with (1×) The samples were diluted and retested. "If the cell culture supernatant sample needs to be diluted in a distributed manner, cell culture medium may be used for intermediate dilutions, except for the last step when diluent is used."
4. The difference of the test results can be caused by a variety of factors, including the operation of the experimenter, the use of the pipettor, the washing technique, the reaction time or temperature, the storage of the kit, etc.
5. The termination solution in the kit is acidic. Please protect your glasses, hands, face and clothes when using it.
6. For scientific research only, not for in vitro diagnosis.
Storage Temp. Unopened kit, 2-8° C Storage
Test Range 62.5pg/mL-4000pg/mL
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SKU: 22827410119

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❈ Elizabeth ❈ | Breakawayreads
Phoenix, US
★★★★★ 5
Fallen Angels, fae, vampires, oh my!
Format: Kindle
Rating: 4.5 | Spice: 2 (but a good slow-burn) • Main Characters: Huntyr and Wolf • I couldn’t wait to read this book; there was so much hype about it! And there was no doubt why. I fell in love with the characters and the plot itself. This book is mainly plot driven more than friction driven but it’s easy to follow along with. The characters are fun, easily understood. The main setting is at an academy where both the main characters are going through trials and building strength for the final test, The Transcendent. There are fantastic side characters as well. I loved the camaraderie between Huntyr and her friends. But we don’t like Lanson. 😆 We do have some plot twists that come into play throughout the book. Secrets and betrayal to be seen. I did adore Wolf and Huntyr’s relationship. It was a classic slow burn trope. They didn’t hit it off fast, but in time their feelings grew. I loved their banter, so sexy. Wolf is your next book boyfriend; Huntyr is your next vampire assassin independent bad-a*s female. Themes include loyalty, trust, self-discovery, a true slow burn romance. Side note: book ends on a angsty cliffhanger! • Emily, thank you for writing this awesome novel and I cannot wait to devour Book 2, Blood So Brutal! 😍 • Happy reading, my lovelies! xo
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Reviewed in the United States on January 21, 2024
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MelsABookworm
Draper, US
★★★★★ 4
“My heart bows to you and you only, Huntress.”
Format: Kindle
3.5 🌟 This book popped up in my KU recommended reading suggestions and the synopsis sounded like what I was in the mood for. I'm so glad I took a chance on it. I went into this knowing absolutely nothing about it and ended up really liking it. I love when this happens. The main characters are likeable and I easily found myself rooting for them. There is a mystery element to each of their backstories that I enjoyed watching unfold and can't wait to get more of. Wolf, in particular, has me fixated. Love him. I found this to be an entertaining, addictive read with a plot that moves along at a good pace. It reads so easily I found myself very reluctant to put it down. Lots of twists and turns and the angst is there. A good set up for the next book to come, for sure. My issues with this book....the dialogue feels a bit juvenile at times and there is a repetitive over use of a particular word phrasing that I found myself giving the ole eye-roll to. There are, without a doubt, some pretty cliche moments that gave me a bit of the cringe. I think this could've certainly 100% benefited from more depth regarding the world building. Perhaps the world building was sacrificed to keep the pacing quick? Just a guess. Also, the lack of consistency of character for the FMC was really evident and so she feels quite illogical at times. Overall, this was a fun and enjoyable read that hit the spot well enough for me. That ending certainly has me impatiently pining for book 2!
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Reviewed in the United States on June 18, 2024
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Amazon Customer
Boise, US
★★★★★ 3
Interesting take on the genre
Format: Kindle
True rating: 3.25 ⭐️ I enjoyed the fresh take on the genre. The best way I could describe the setting and world is an apocalyptic dystopian version of Farie where vampires, fae, and angles struggle to survive in what is left of the world. It was definitely interesting throwing the academy/hunger games aspect into this world as well. Even though I guessed the final reveal early on in the book, I kept hoping I was wrong, and it would take a surprising turn. While the "plot twists" were a bit predictable to me, I still enjoyed the ride this book took me on. Another downfall for me was the plot holes in the world building... I.E. if society has fallen and the world is in the aftermath of war, how are there trains running around the world? Just to take young adults to the trials to get into the golden city? How is the train maintained, the tracks clear, etc? However, I did enjoy the FMC & MMC and thought they were fleshed out nicely. I also enjoyed the side characters but wish some were developed more like Ashalin (sp?). I do find myself rooting for the MCs to succeed and find happiness together, which is obviously an important aspect for romantasy. Overall, was this an earth-shattering, mind-bending, terrific piece of literature? No. But was it the worst thing I've read this year? Also, no. This book has, to me, the bones of a great read & just needs a bit more to push it from an alright book to a great book. Overall ratings: Plot- 3.5⭐️ World building 3⭐️ Spice 2.5 🌶🌶 Main characters 4 ⭐️ Supporting characters 3.5⭐️
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Reviewed in the United States on May 12, 2024
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Irene zamora
Cuba, US
★★★★★ 5
great book
Format: Kindle
I am really excited to meet the author at the book retreat this month. I really enjoyed this world that she built and most of the female main character Huntress is so awesome. She goes through a lot in this book and the ending; wow! I wouldn't have even guessed. I highly recommend everyone to read this book. I have been so lucky this year that almost all the books I have read have been, so far, 5 out 5 stars.
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Reviewed in the United States on June 2, 2026
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Anastasia Goygova
Pawtucket, US
★★★★★ 4
Fallen for the Fallen Angel – A Guilty Pleasure Worth Every Page
Format: Kindle
There’s something deeply irresistible about a dark academia or trial-based setting, a brooding and arrogant fallen angel, and a fierce heroine with enough sass to go toe-to-toe with him. Wings So Wicked is exactly that kind of book—and I devoured it in just a couple of days. To be fair, the plot isn’t groundbreaking. If you’re looking for something fresh and innovative in terms of storyline, this might not be it. But if your reader heart beats faster at the mere mention of enemies-to-lovers, jealousy-fueled banter, magical trials, betrayals, and forbidden tension—you’ll feel right at home. It’s like catnip for those of us with this particular weakness. The chemistry between the leads could have used a slightly slower burn to make the tension sizzle longer, but I still found myself completely invested in their dynamic. There are moments and phrases that feel a bit cheesy or underdeveloped, but honestly? I didn’t care. The vibes were exactly what I wanted. This book isn’t trying to reinvent the genre—it’s here to give readers like me what we crave: high-stakes magical drama, angsty romance, and the thrill of watching a badass girl and her brooding counterpart clash and spark. If that sounds like your kind of story, Wings So Wicked will hit the mark. Here’s hoping Book 2 turns up the heat and keeps the magic alive.
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Reviewed in the United States on May 20, 2025

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