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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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4.7 ★★★★★
Based on 2087 reviews
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Product Reviews
★★★★★ 2
Sad
Pattern Name: Strawberry
The outer part is good quality, the removable inside didn't last a day
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 29, 2026
★★★★★ 3
Didn’t hold up for more than a minute but the silicone strawberry by itself is durable.
Pattern Name: Strawberry, Pattern Name: Strawberry
Our best dog toy reviewer, a cattle dog mix, gave this toy a big thumbs up ONLY because she could demolish it in seconds. Not minutes, but seconds. The toy has a pouch stuffed into a silicone toy strawberry. The pouch is really stuffed in well, so much so that I couldn’t remove it, but she sure did! In about a minute all that was left was the dismembered strawberry “leaves” and a silicone strawberry which, oddly enough, she seems to enjoy. The silicone strawberry seems quite durable by itself. The toy is brightly colored and small in size.
This not a durable toy unless you have a dog that is not an aggressive chewer, but for us, it is not recommended.
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Reviewed in the United States on March 15, 2026
★★★★★ 4
Good for larger treats, not small ones
Pattern Name: Carrot, Pattern Name: Carrot
Our dog loves this treat toy. It has been very durable as far as the rubber part goes but the cloth carrot insert not so much, that got torn apart pretty quickly. Also this works for bigger treats, but once the cloth carrot was destroyed, there was no way to keep small treats from just falling out.
Now we use it to stick larger chew sticks in the holes and it works to help her keep it in place while chewing.
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Reviewed in the United States on May 11, 2026
★★★★★ 5
Dogs love these treat toys
Pattern Name: Strawberry, Pattern Name: Strawberry
I have big dogs who live for treats and love to play puzzle games to find treats. You can stuff these toys with small treats which gives your dog a mental challenge while working to figure out how to get them out. The quality of these toys are great but durability with big dogs will be short lived. The strawberry green top immediately was de-stuffed while dog was looking for more hidden treats. The corn husk seems to be more challenging for them to get the treats out. However my smarter dog was determined and eventually reaped the rewards. I would recommend these toys for dogs who will not rip them apart.... but really is there any pup who doesn't enjoy de-stuffing a stuffie squeek toy?
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Reviewed in the United States on May 14, 2026
★★★★★ 5
Great feeder for high energy puppy!
Size: Large, Color: Blue
I have a 50lb high energy puppy. She is not a fast eater, BUT she loves this activity and it gives me about 30 minutes of uninterrupted bliss. It took about 10 minutes to demonstrate to help her get the hang of it. She will sit, spin the barrel with her paw, then sniff every divider, only open the ones with her food, and then restart the process. It is sturdy and stays on the floor as she paws and pushes. It might move a little, but not far. This is my favorite puzzle toy by far and I love that it is "hard" enough to keep her occupied but not enough that she gets frustrated and doesn't want to do it.
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Reviewed in the United States on June 7, 2026