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Description
Rat LTb 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 specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 3. 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 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 1000pg/mL). Then dilute to the following concentrations: 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.625pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 1000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 500pg/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 a lymphotoxin beta (LTb) 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 lymphotoxin beta (LTb) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Lymphotoxin Beta ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Lymphotoxin-β (LTb), also known as tumor necrosis factor C (TNC), is a protein encoded by the LTB gene. It is a type II membrane protein of the TNF family. It anchors lymphotoxin-α to the cell surface through heterotrimeric formation. The predominant form on the lymphocyte surface is the lymphotoxin-α/β2 complex, which is the primary ligand for the LTb receptor. It is an inducer of the inflammatory response system and participates in the normal development of lymphoid tissues. The b isoform is unable to complex with lymphotoxin-α, indicating that its function is independent of lymphotoxin-α. Its expression influences immune responsiveness at two distinct levels: its expression on lymphocytes enhances the induction of immune responses to limiting amounts of antigen, while its expression on non-lymphocytes controls antiviral immunity by enhancing antigen presentation on antigen-presenting cells. This prevents exhaustion of cytotoxic T lymphocytes or host death due to uncontrolled viral spread. | |||||||||||||||||||||||||||||||||
| 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 | 15.62-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.1 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 4
*update* Maybe(?) Perfect for those who hate eggs
Edit: Update
It is with a heavy heart Im updating this review. When I said this sponge stains, I mean it really stains. Now to be fair it is marketed as a powder sponge and I use it with liquid so this my be entirely on the user (aka me) but it's like I can never get it clean. Between my repeated washing and the overall product build up the performance of this sponge went downhill quickly. The finish of my makeup suffered and the flocking took a beating. For what it's worth the sponge has not fallen apart and is still in one piece. But I get more of a 4 star finish than a 5 star finish.
I've ended up using a large fluffy brush to buff out my foundation to get the desired result. I also ended up repurchasing the flocked flat ended egg from RT to replace this although at the moment I am still stubbornly using this sponge (and also avoiding doing my makeup)
I do believe a lot of my issues are do to using this sponge with liquid foundation instead of power and that is on me. I'm finding it so very difficult to clean and the RT flocked egg cleans up so much easier. I don't hate this sponge but my lovefest has definitely taken a turn to more of a likefest. I am going to keep using (because I am stubborn like that) and see how long it takes to turn this relationship into a hatefest or fall apart completely, whichever happens first. I will most likely update again at that time. At this moment I am changing my review from 5 star to 4 star. I will be keeping my original review below.
*original review*
Yo I LOVE this thing. Ok here's the deal, I don't like the beauty blender, in fact I kinda hate it. Yea, I said it, I'm not on board with bouncing an egg all over my face, I think it's tedious and inefficient.
I had at one time a flat teardrop shaped sponge that was my hands down favorite application tool for foundation. I used that thing until it literally fell apart. I tried to find a replacement but the store I had originally gotten it from no longer carried it so I kept using that ratty old sponge until it was missing chunks.
I tried some other flat sponges but none worked as I wanted so I went back to brushes but I really hate cleaning foundation from brushes so I was pretty much hating my life.
Then I found a flocked egg. Now I'm not a fan of the eggs but I was intrigued by the flocked texture and it had a nice flat edge so I tried it and it was almost as good as my old dearly missed flat teardrop sponge. I still was not a fan of bouncing an egg around on my face but I used this until the flocking wore off (about 6 months)
Well, I saw this listing for a flat flocked sponge and I pounced on it. I am so incredibly deliriously happy with this little sponge. I love that it's flat and not an egg. I love the flocking. I love how it fits in my hand. I love love the finish I get with my foundation. I love it. Like I need to order a dozen more so I never run out and I'm trying to break my hoarding tendencies but for this tool I want to hoard.
So the bad... It does stain easily. I wet my sponge before use and give it a quick clean after use. My old flocked egg would clean right up but this has stained on the first use. I also don't know how long this will hold up so I hope to update at a later time about that.
Overall I couldn't be happier with this sponge.
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Reviewed in the United States on February 10, 2021
★★★★★ 5
Perfect for liquid
This sponge is not made for liquid but I use it for my liquid foundation. The sponge won't last as long when it gets wet but it still lasts a LONG time! I put a drop of water on sponge first then dab a few times on my arm to dry with only a slight hint of moisture left (very dry feeling)... then add a drop of makeup (literally). The coverage is awesome and your makeup last 2 to 3x as long. Flawless finish. Love it!
I recently updated my review...I still have same awesome sponge. Now I also mix a silicone air brush liquid (clear) to the liquid makeup it spreads farther and the finish is wonderful. Great product stays dry and drys very quick after use.
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Reviewed in the United States on January 19, 2014
★★★★★ 5
Love this product!
I’ve used this product for a long time. It’s nice and soft and works well on my skin. I wouldn’t use anything else.
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Reviewed in the United States on July 29, 2024
★★★★★ 3
Its OK, but not nearly as easy to use as videos and reviews make it seem.
I ordered this flocked sponge based on reviews and on watching JI videos of applying pure pressed powder with it. I tried it 'damp' and it was awful mess! I tried it dry, and no powder was transferring to my face at all. I'll stick with my ELF blurring brush to apply JI pressed foundation (which I love).
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Reviewed in the United States on October 12, 2018
★★★★★ 5
Jane Iredale sponge applicator
I’ve bought this for years and I find there the easiest and best way to even out your make up. I like them better than brushes.
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Reviewed in the United States on October 25, 2025