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Description
Mouse TMX1 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: 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: 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. Plasma: Collect specimens using EDTA or heparin as an anticoagulant and centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-Test Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 ng/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 uses 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 capture antibody against thioredoxin-related transmembrane protein 1 (TMX1). 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 thioredoxin-related transmembrane protein 1 (TMX1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Thioredoxin-related transmembrane protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Thioredoxin-related transmembrane protein 1 (TMX1) is a protein encoded by the TMX1 gene. TMX1-associated disorders include ciliary dyskinesia, primary, and 6. Gene Ontology (GO) annotations associated with this gene include protein disulfide isomerase activity and disulfide oxidoreductase activity. An important homologue of this gene is TMX4. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, cell culture supernatant |
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4.3 ★★★★★
Based on 7 reviews
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Product Reviews
★★★★★ 5
Ends in a cliffhanger
Format: Kindle
Thankfully, the next book is already available.
Roxy Collins is the queen of omegaverse slow burn. This particular series is omegaverse with wolf shifters.
Thoroughly enjoyed this book! Lots of suspense and trying to figure out what is true and what's really going on. I loved it!
Pretty spicy once the action starts. Heats are pretty much always spicy. There is a little bit of male interaction, but not full-on shmex.
Definitely recommend.
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Reviewed in the United States on April 29, 2025
★★★★★ 5
Epic!!!!
Format: Kindle
Absolutely addicted from the first page. The twists and turns, world building and character development all come together for the perfect Omegaverse. And let's not forget that spice because knots are life and these are spectacular. Running for book #2 because that cliffhanger was torturous.
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Reviewed in the United States on August 17, 2024
★★★★★ 5
Wow.
Format: Kindle
Okay I’m usually not one for stand-alone’s I’m an avid series reader but my goodness am I so happy I read this! This story was brilliant & so absolutely mesmerizing. I loved reading about each character and their struggles as well as what helped them to move forward. The ending definitely brought tears to my eyes so hard. I truly wasn’t expecting some of what happened in this story. There is about to be a spoiler I am going to reveal so please stop reading if you don’t want the spoiler !!!! ⚠️ ⛔️ ‼️
I loved that the author didn’t do what most authors do with irredeemable male characters. I truly was hoping that Nate Jr. would be apart of the pack after the way he treated Astrea bc he truly didn’t deserve it. Though I must say you did a wonderful job or redeeming him as a person. I cried my eyes out when he walked into the story. I was truly terrified he was going to be a bad guy to the end. However you truly did him such a justice by having him realize his faults & having him redeem himself in the most wonderful way. I’m so sad that he didn’t get to hear how much his brother loved him & forgave him before dying. But again you wrote that ending so beautifully & I just can’t express how much I loved this story & how you took a different route than most authors I have read have. You are a remarkable author Cinder Blaze & I thank you generously for creating such a masterpiece.
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Reviewed in the United States on April 28, 2025
★★★★★ 4
Omegaverse
Format: Kindle
I was pleasantly surprised by this omega verse book. This 1 definitely had a few new twists And I really enjoyed reading it. The main female character was a badass and awesome. She had 1 best friend I wish we could have seen a little bit more of their friendship in this book. Her relationship with the male characters was good not to contrived or super instantaneous. And we had some fun plot twists that I didn't expect. I wish we had more of a follow up on the situation with her family her background and her mother who was a wench. I would definitely recommend this book.
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Reviewed in the United States on April 9, 2025
★★★★★ 5
Wow. Just wow. An amazing read
Format: Kindle
This book was eloquently written, or should I say the authors writing style is very eloquent? I loved the characters, and the story was quite compelling. I absolutely loved the FMC, she's a BA who doesn't take any crap & gives as good as she gets. A certain person certainly got his just desserts & then some, the earlier scenes of which were quite satisfying, had me punching the air & everything haha. All in all 20/10, great read
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Reviewed in the United States on November 29, 2024