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Description
Mouse TNF-α ELISA KitProduct Specification protein TNF Usage Need to bring your own test equipment 1. Microplate reader (measurable 450nm Absorption value of detection wavelength and 540nm Or 570nm Absorption value of corrected wavelength) 2. High precision liquid dispenser and disposable tip 3. Distilled or deionized water 4. Bottle washer (spray bottle), multi channel plate washer or automatic plate washer 5. 500mL Measuring cylinder 1. Preparation before the experiment
Product Specification
| protein | TNF-α | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Usage |
Need to bring your own test equipment 1. Microplate reader (measurable 450nm Absorption value of detection wavelength and 540nm Or 570nm Absorption value of corrected wavelength) 2. High precision liquid dispenser and disposable tip 3. Distilled or deionized water 4. Bottle washer (spray bottle), multi-channel plate washer or automatic plate washer 5. 500mL Measuring cylinder 1. Preparation before the experiment 1. Sample collection and storage ① Cell culture supernatant: particulate matter should be removed by centrifugation; Test the sample immediately. If the sample is not tested in time after collection, it is recommended to pack it according to the amount used once and store it frozen in -20℃ In the refrigerator, avoid repeated freezing and thawing. The sample may need to be used with a diluent ( 1× ) dilution. ② Serum: Use serum separation tubes ( SST ) Collect samples and place samples at room temperature 30 Minutes. Centrifugation 15 Minutes, with a rotation speed of 1000g 。 The serum was removed immediately and tested immediately. If the sample is not tested in time after collection, it is recommended to pack it according to the amount used once and store it frozen in ≤-20℃ In the refrigerator, avoid repeated freezing and thawing. The sample may need to be used with a diluent ( 1× ) dilution. ③ Plasma: Use EDTA , heparin or citric acid as an anticoagulant to collect plasma, after collection 30 Centrifuge within minutes 15 Minutes, with a rotation speed of 1000g , and detect it immediately. If the sample is not tested in time after collection, it is recommended to pack it according to the amount used once and store it frozen in ≤-20℃ In the refrigerator, avoid repeated freezing and thawing. The sample may need to be used with a diluent ( 1× ) dilution. 2. Reagent Preparation ( Please place all reagents and samples at room temperature before use and let them stand 15 Minutes. All experimental samples and standards are recommended Do repeat hole detection ） ①1× Preparation of washing solution: The concentrated washing solution in the kit is 20× Mother liquor should be diluted with distilled water before use to 1× Working fluid. Example: Take 10mL Concentrated wash +190mL Distilled water to volume to 200mL In actual operation, the usage amount can be calculated first, and then prepared. ②1× Preparation of buffer for dilution: The concentration and dilution buffer in the kit is 10× Mother liquor, dilute with distilled water before use to 1× Working fluid. Example: Take 3mL Buffer for concentration and dilution +27mL Distilled water to volume to 30mL 。 In actual operation, the required amount of dilution buffer solution can be calculated according to the sample dilution factor, and then prepared. ③ Antibody detection: Centrifuge the dry powder to the bottom of the tube and use 110uL Buffer for dilution ( 1× ) Dissolve and let stand at room temperature 5 Get after minutes 100× Mother liquor; Before use, dilute to 1× Working fluid. According to the amount per well 100uL Calculate the required volume. Example: Used 10 Hole, then take 10uL Of 100 Double working concentration of detection antibody, using dilution buffer ( 1× ) Constant volume to 1mL , obtained 1mL Of 1× Detection antibody at working concentration. ④SA-HRP ： SA-HRP For 40× Mother liquor, use dilution buffer before use ( 1× ) Diluted and formulated 1× Working fluid, the required amount per hole is 100uL 。 Example: Used 10 Hole, then take 25uL Of 40× Mother liquor +975uL Buffer for dilution ( 1× ) Constant volume to 1mL , obtained 1mL Of 1× Detection antibody at working concentration. ⑤ Developer: per well 100uL Calculate the dosage required for the current test, take out the corresponding volume of color developer, and protect it from light; The developer removed is for the same day use only. ⑥ Standard: Dilution buffer for lyophilized standard ( 1× ) Re-dissolved, redissolved volume 1000uL , obtaining a concentration of 2000pg/mL Standard mother liquor. Gently shake at least 5 Minutes, it is fully dissolved. Add to each dilution tube 300uL Buffer for dilution ( 1× ）。 Make serial dilutions of the standard mother liquor according to the figure below, and each tube must be thoroughly mixed before pipetting to the next tube. Standard mother liquor without dilution can be used as the highest point of the standard curve ( 2000pg/mL ), buffer for dilution ( 1× ) can be used as a standard curve zero ( 0pg/mL ）。 2. Operation steps 1. Prepare all required reagents and standards; 2. Take out the microplate from the sealed bag that has been equilibrated to room temperature. Please put the unused slats back into the aluminum foil bag and re-seal; 3. Add to the microplate 300uL Washing liquid, let stand and soak 30 Seconds, discard the lotion and pat the microplate dry on absorbent paper. Please use it immediately and do not let the microplate dry; 4. Add different concentration standards, experimental samples or quality control articles into the corresponding wells, and each well 100uL 。 Seal the reaction wells with plate sealing tape and incubate at room temperature 2 hour ; 5. The liquid in the plate is sucked off and the plate is washed using a washing bottle, a multi-channel plate washer or an automatic plate washer. Washing solution per well 300uL Then the intra-plate wash liquid is aspirated. Repeat Operation 3 Times. Trying to absorb the residual liquid as much as possible every time you wash the plate will help to get good experimental results. At the end of the last plate washing, please suck all the liquid in the plate or turn the plate upside down, and pat all the residual liquid dry on absorbent paper; 6. Within each well added 100uL Detect antibodies. Seal the reaction wells with plate sealing tape and incubate at room temperature 2 Hour; 7. Repeat th 5 Step washing operation; 8. Within each well added 100uLSA-HRP , room temperature incubation 20 Minutes. Be careful to avoid light; 9. Repeat th 5 Step washing operation; 10. Within each well added 100uL Chromogenic solution, incubate at room temperature 5-30 Minutes, pay attention to avoiding light; 11. Within each well added 50uL Stop solution, the color of the solution in the well will change from blue to yellow. If the color of the solution changes to green or the color changes are inconsistent, pat the microplate gently to mix the solution evenly; 12. After addition of stop solution 30 Within minutes, measured using a plate reader 450nm Absorbance value, set 540nm Or 570nm As a correction wavelength. If dual-wavelength correction is not used, the accuracy of the results may be affected; 13. Calculation results: Add the corrected absorbance values of each standard and sample (OD450-OD540 Or OD570) , average of repeated well readings and then subtract the average zero standard OD Value. Using computer software for four-parameter logic (4-PL) Curve fitting creates a standard curve. Another way is to plot the standard concentration and make the logarithm with the corresponding OD The values were logarithmic to generate a curve, and the best fit line was determined by regression analysis. This process can generate a data fit that is sufficiently useful but less accurate. If the sample is diluted, the concentration should be calculated by multiplying the dilution factor. Note: The standard curve data provided is for reference only, and the sample content should be calculated according to the standard curve drawn in the same test. 3. Kit parameters 1. Recovery: Different levels of mice were spiked in cell culture medium samples TNF-α The recovery rate was determined. The recoveries range from 88-107% , the average recovery was in 94% 。 2. Sensitivity: Mouse TNF-α The lowest measurable dose ( MDD ) is generally less than 5.5pg/mL 。 The lowest measurable value is determined according to 20 The corresponding concentration is calculated by adding two standard deviations to the mean value of the zero-point absorbance values of each standard curve. 3. Correction: This ELISA High purity recombinant mice expressed by Escherichia coli TNF-α Corrected by protein. 4. Linearity: 4 High concentrations of mice were spiked into different samples TNF-α , followed by a diluent ( 1× ) Dilute the sample to the detection range and determine its linearity.
5. Specificity: This ELISA Method can detect natural and recombinant mice TNF-α Egg whites. The following factors were mixed with diluent ( 1× ) formulated into 50ng/mL Concentration to detect with mice TNF-α Cross-reactivity of. Will 50ng/mL Interfering factors incorporated into the intermediate range of recombinant mice TNF-α In the control article, to detect the effect on mice TNF-α Of interference. No significant cross-reactivity or interference was observed.
4. Analysis of frequently asked questions 1. Whiteboard (no color appears after color rendering is complete)
2. Flower plate (blank, negative positive control normal, but specimen well OD Values are significantly higher)
5. Experimental flow chart |
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| Species Reactivity | Mouse | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Theory | This kit adopts double antibody sandwich enzyme-linked immunosorbent detection technology. Specific anti-mouse TNF-α antibodies were pre-coated on high affinity enzyme labeled plates. The standard substance, the sample to be tested and the biotinylated detection antibody are added to the well of the enzyme label plate, and after incubation, TNF-α present in the sample binds to the solid phase antibody and the detection antibody to form an immune complex. After washing to remove unbound material, horseradish peroxidase-labeled Streptavidin-HRP was added. After washing, a chromogenic substrate is added to protect the color from light. A stop solution was added to stop the reaction, and the absorbance value was measured at a wavelength of 450 nm (reference calibration wavelength of 540 nm or 570 nm). | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Synonym | APC1 protein, Cachectin, Cachetin, DIF, TNF, TNF, monocyte-derived, TNFA, TNF-A, TNFalpha, TNF-alpha, TNF-alphacachectin, TNFATNF, macrophage-derived, TNFG1F, TNFSF1A, TNFSF2, TNFSF2TNF superfamily, member 2, tumor necrosis factor ligand superfamily member 2, tumor necrosis factor, tumor necrosis factor-alpha, Mouse tumor necrosis factor alpha ELisa kit | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Wells | 0 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Composition |
Please use it within the validity period of the kit (new and old products are shipped randomly)
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| Background | Tumor necrosis factor alpha (TNF-α), also known as cachectin and TNFSF1A, is the prototype ligand of tumor necrosis factor superfamily. It is a pleiotropic factor that plays a central role in inflammatory response, immune system development, apoptosis and lipid metabolism. TNF-α is also involved in many pathological processes, including asthma, Crohn's disease, rheumatoid arthritis, neuropathic pain, obesity, type II diabetes, septic shock, autoimmunity, and cancer. Human TNF-α is a 26kDa type II transmembrane protein consisting of a 35 amino acid (aa) intracellular domain, a 21aa transmembrane segment, and a 177aa extracellular domain (ECD). In the ECD region, human TNF-α has 97% amino acid sequence homology with macaque monkey, and 71-92% amino acid sequence homology with cattle, dog, cotton rat, horse, cat, mouse, pig and rat. It can be expressed by a variety of different cells such as immune cells, epithelial cells, endothelial cells, tumor cells. TNF-α can induce lysis of tumor cells and virus-infected cells, and binds to soluble TNFRI to generate transmembrane transfer signals of downstream cells. TACE/ADAM17 can cause the shedding of TNF-α-containing cell membrane to release active TNF-α cytokine. It is a trimer composed of TNF-α extracellular soluble structure with a molecular weight of 55 kDa. TNF-α has two receptors: TNFRI and TNFRII. TNFRI has a molecular weight of 55-60kDa and is widely expressed; TNFRII has a molecular weight of 78-80 kDa and is limited to hematopoietic cell expression. Both are expressed as homotrimers; TNF-α and TNFRI and TNFRII bind with similar affinities and can promote the activation of NFκB. However, only TNFRI has a cell death domain, which can trigger apoptosis. Both soluble receptors are released into human serum and urine and can neutralize the activity of TNF-α. |
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| General Notes | 1. Please use the kit within the validity period. 2. The components of different kits and different batch kits cannot be mixed. 3. If the sample value is greater than the highest value of the standard curve, the sample should be diluted with diluent (1 ×) and re-tested; If the cell culture supernatant sample needs to be distributed and diluted, cell culture medium can be used for other intermediate dilutions except dilution with diluent in the last step. 4. Differences in test results can be caused by a variety of factors, including the operation of the experimenter, the use of the pipette, the plate washing technique, the reaction time or temperature, the storage of the kit, etc. 5. The terminating solution in the kit is an acidic solution. Please protect your glasses, hands, face and clothes when using it. 6. For scientific research only, not for in vitro diagnosis. |
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| Storage Temp. | Kit unopened, stored at 2-8 °C. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Test Range | 31.3pg/mL-2000pg/mL |
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4.6 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 5
Very nice pillow
Color: Cut-out White, Size: King
I was skeptical of spending this much money on a pillow, but so far I'm glad I did.
It's very comfortable, and the shape does seem to help. I injured my neck/shoulder years ago, and I would wake up 1-2x per week heading slept wrong making my neck and shoulder stiff and painful in the morning. This pillow has so far prevented that.
This pillow has also significantly reduced my snoring, according to both my Samsung watch and my wife.
I'm a side sleeper.
This pillow can be as firm or soft as you want since you can add or remove filling. It comes with a bag of extra filling just for that.
I wouldn't say its cooling, but it certainly isnt too warm either. And I'm someone who sleeps hot.
All in all, I'd buy it again.
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Reviewed in the United States on May 28, 2026
★★★★★ 5
this is it (hopefully), no chemical odor
Color: Original White, Size: Queen
Over the years, I have spent a lot of time and money buying pillows. I can never seem to find one that fits my needs. I thought this would be a good option to try because this came up as a top rated pillow from consumer reports.
I really think this pillow might be the last pillow I ever need. I saw all the good reviews and I also read all the bad reviews. Pillows are very personal, and a pillow that works for one person may not work for the other person but, I decided to give this a try.
I was so excited. This was on sale at Amazon and I got it same day. I have a lot of neck issues and pinched nerves so I thought this pillow would help. Once I took it out of the box, I fluffed it up by hand because I don’t have a dryer. Within 15 minutes the pillow was really fluffy. I tested it out. I’m a side sleeper. It didn’t work too well. I removed some of the stuffing which is luxuriously soft. And it seemed to be comfortable. This morning I woke up with a horrible neck and headache. So, I took a little more stuffing out. It seems to feel better now. This gives just the right amount of support. It doesn’t give too much pushback and it also doesn’t go flat. Yes, this will flatten out a little bit when you lay your head on it, but not completely to the mattress flat. I fluff it up before I go to bed and it works great. I think I have it at just the right thickness but we will see how it goes. Surprisingly, it was not too hot for me like others have reported. I do have a temperature control system on the bed so maybe that’s why the pillow temperature didn’t bother me as much. I did not have any pain in my ears from sleeping on a hard pillow. That was a big plus. It gives Justin enough support and not too much pressure so it’s perfect. The outer shell is really nice. The shell and zippers seem to be good quality. I still use a genuine silk pillowcase because I like it. I think this is my forever pillow. I will add an update after I use it for a while if anything changes. I am hoping for good durability. Last thing, this also comes with extra stuffing if you like your pillow, extra high, or if your pillow starts to go flat, you have more stuffing to put in it. Putting your pillow out in the sun or in the dryer once every couple of months will help to keep it fluffy. I don’t think I would ever wash the foam on the inside. There’s no reason for that. The outer shell is what’s going to get most dirty.
Just know, I liked it so much I bought another one immediately.
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Reviewed in the United States on November 21, 2023
★★★★★ 4
Rediscoving the Great Qualities of this Pillow (after Originally Rejecting It)
Color: Original White, Size: Queen
I recently 'rediscovered' this pillow... having purchased it a year or so ago, when neck + shoulder issues prevented me from being very happy with *ANY* pillow! Believe me, I tried so many! Back then, after repeatedly adjusting the fill of this COOP queen-size pillow, I ultimately stashed the whole thing in a closet. But now, it has unexpectedly re-entered my life... and I'm finally liking it.
It helps that I've resolved most of the neck + shoulder issues (thanks to a year of PT). Plus. I realized that I had removed way too much fill... leaving about as much support as a pile of gently scrambled eggs. So, now that I'm in much better condition and have added back a lot of fill, I'm finding this pillow quite satisfying... yes, actually good! These days, I wake up each morning with a much happier neck.
I do not. however, use the thick, quilted, zippered outer cover that came with it... as I find that stiffens + firms up the pillow a little too much for my taste. So, on top of the thinner, stretchy, zippered inner cover (also provided with the pillow, to contain the fill,) I've replaced the quilted outer cover with a jersey-stretch, zippered, cotton pillow protector. which is lighter weight and more malleable.
As a person who has been traveling for years—by necessity—with a personal pillow from home, I'm now recognizing the extra advantages of this COOP pillow: For travel, its parts all compress well + easily (while for sleeping, the fill is light + lofty, but also supportive, as long as you don't remove way too much). And the pillow's components all readily come apart, so they can be packed separately...even the stuffing can be separated into smaller zip-lock packets, if needed—meaning there's never a need to squish an entire, impossibly big, somewhat dense pillow into a suitcase (or even a compression sack). Quick + easy to disassemble and reassemble. Really, the COOP can be treated as a build-your-own pillow (remember, I changed out the zippered outer cover and adjusted the stuffing volume to my own prefences). It's not cheap, but these days, remarkably, it's doing the trick for me. So glad I didn't get rid of it.
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Reviewed in the United States on February 27, 2026
★★★★★ 5
Great pillow and company
Color: Crescent White, Size: King
I have a couple of Coop Home pillows and loved them. They are several years old and I was on the search for a replacement. I tried 2 other brands, one cost more and the other cost less. Returned 1 and used the other for a short time before I had to go back to my old Coop pillow. I started looking at crescent pillows and did some research. This type of pillow is great for side and back sleepers. Being a side sleeper I looked into the best rated. Coop Home crescent pillow was not rated #1 but it was close. The top rated pillow cost 2.5 times more and I could not justify paying that. I am happy to report that I am waking up less (before every 1.5-2 hours). It comes with extra fill and I love a very firm pillow so I emailed customer service to see if I could get more filling. This was on the weekend and in 1 day they responded that they would be happy to. Now that is great customer service. I sleep warm and have no problem with a hot head. It comes with a cover but I use silk pillowcases and it works fine. I will say when I first got the pillow it was very thin and flimsy. I had some doubts but followed directions and put in the dryer on low. Came out perfect. Highly recommend this pillow and company.
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Reviewed in the United States on April 20, 2026
★★★★★ 5
Best pillow I've ever owned
Color: Cut-out White, Size: Queen
While the pillow looks anemic when it first comes out of its packaging, it puffs/firms up beautifully after time in the dryer. I haven't had any odor issues.
I'm mostly a side sleeper and at first I thought there wasn't enough of the extra fill, so I put it all in. I've ended up needing to take at least half back out to keep my head at the correct angle.
While this can fit in a standard pillow case, it just barely fits and keeps coming out - at least at my fill level. Queen pillow cases are much better.
This is so much more comfortable than any of my previous pillows. I don't need to stack multiple pillows to get the right support. I'm no longer waking up with neck or shoulder or arm pain. I'm recommending it to pretty much everyone and I've purchased a second pillow to keep at my parents for when I visit!
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Reviewed in the United States on May 6, 2026