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Description
Rat TFF1 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. Preparation of 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 5000pg/mL). Then dilute to the following concentrations: 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312.5pg/mL, 156.25pg/mL, 78.125pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 5000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 2500pg/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 Trefoil Factor 1 (TFF1) 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 Trefoil Factor 1 (TFF1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Trefoil Factor 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Trefoil factor 1, also known as TFF1, is a protein encoded by the TFF1 gene (also known as the PS2 gene). It is a member of the trefoil family, characterized by at least one copy of the trefoil motif, a 40-amino acid domain containing three conserved disulfide residues. They are stable, secreted proteins expressed in the gastrointestinal mucosa. Their functions are not yet defined, but they may protect the mucosa from insults, stabilize the mucus layer, and influence epithelial healing. This gene and two other related trefoil family members are found in a cluster on chromosome 21. Its expression is frequently lost in gastric cancer, possibly through DNA methylation, and it is therefore considered a tumor suppressor gene. It is primarily expressed in the epithelial surface cells of the fovea of the gastric mucosa and exhibits an increased expression pattern during inflammation. It and other TFF family members are upregulated and secreted in an autocrine and/or paracrine manner in response to inflammation and gastrointestinal injury. The signaling pathways it activates are not fully elucidated, primarily due to a lack of data identifying its functional receptors. | |||||||||||||||||||||||||||||||||
| 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 | 78.12-5000pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.0 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 5
One of the greatest cookbooks I’ve ever bought
Format: Hardcover
I love cookbooks—like really really love them. I have a collection. I just finished reading this through on the night I received it via preorder—I tracked it all day because I was so excited. I just intuitively felt like after looking at the images and preview that there was really something special about this one. Being a cookbook lover, you can just smell it. This one seemed dense with intention and value, and so colorful, accessible and of-the-moment. And boy was I right!! While there literally hasn’t been enough time to cook anything from this yet (it arrived on my door step three hours ago), I can already tell you that this thing has more soul and voice than 99% of the cookbooks out there. She had me LAUGHING out loud like 12 times no exaggeration (Maldon in the purse?! F yeah ur my girl!), misty eyed (last meal convo), IMPRESSED (QR code for real time audio instructions w classical music?! and vids?! #INNOVATION), thrilled at the ingenuity (salt scoville unit rating omfg), and just overall really knocked back by the joy of seeing someone really live their dreams and GIVE IT to the world. It is so palpable that Molly is living her dreams, and you can tell she works for it. Her writing seems more like her true or embodied self than her spoken words in old YouTube vids I recall. This book is aspirational from a lifestyle perspective but it has nothing to do with the very cool graphic design, great photography and food styling (all wonderful) or her ultra hip house or LA social life or whatever… which is all fun to peek in on, but—it’s because this is a person who has found her calling and is LIVING IT 👏👏 … COMMITTING… and generously sharing herself in the most authentic way with us. It’s just so HER, so specific, which is what we wanna see in ARTISTS. And while I’ve followed Molly with appreciation for years (I never got around to buying her first book but live for her gochujang chx and other Bon Appetit recipes), I’m realizing I have either been sleeping on the true depths of her talent OR she has recently bloomed into just like, her fullest self and I am so so happy for her. I came for the ingredient lists organized by grocery store layout (F-ING FINALLY FINALLY THANK YOU THANK YOU) and the fun vintage typography/nostalgia and umami bombs and gorgeous meatballs, but the inspiration that this book vibrates with (GO FOR IT! Life the life you are meant to live!) will stay with me for quite some time. As a former cooking instructor I know the critical importance of building confidence in order to be a good cook but she just took that concept and blew it to its heights/made it REAL. Can’t wait to make cauliflower salad w vegan ranch, leek za, coconut cornbread, fish n shingled potatoes and orange creamsicle poppy cake. THANK YOU MOLLY!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 11, 2023
★★★★★ 5
Molly Snapped. AGAIN
Format: Hardcover
I’ve made a few recipes from this book already and dear god, they’re so good. The hot sauce-braised short ribs and chile-basted half chicken in particular were jaw-droppingly delicious!!
One thing I keep seeing is that she reused recipes from her subscription The Club, so maybe don’t get this book if you already have that. I’m not a member of The Club, so these were all new and exciting for me.
Another thing - this is not a dry, straightforward cookbook. It’s focuses on fun graphic design and has her signature lingo in it. Yes, the font is kooky, and I’m here for all of it. It’s not illegible, but know what you’re signing up for.
And finally, this is not a health cookbook. She has never claimed it is, so I don’t know why some reviews are up in arms about the health aspect. It’s literally called More is More, guys. Some recipes are indulgent, and that’s okay! I try to eat somewhat healthy, and there’s still a lot in it that I could make without modifications.
Basically, if you want a very fun, eye-catching cookbook with DELICIOUS recipes, this is for you!
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Reviewed in the United States on October 31, 2023
★★★★★ 5
Amazing recipes, but a visually exhausting thing to read
Format: Hardcover
The recipes in this book are amazing. Reading through them and flipping back and forth through the pages you can tell Molly put a lot of time and attention into making powerful recipes packed with flavor, with all of them being incredibly approachable in actual technique. There are too many good things to call out with this collection of recipes.
But existing as an easily digestible and referenced cookbook, it kind of sucks.
The editing in this book is supposed to be *fun*. There are random font styles in every page and sometimes there are weird lines that become wavy. There are points in the book where there’s suddenly a cocktail recipe thrown into a chapter about salads or sandwiches. Great sounding cocktails, but ones you’ll need to scan the recipe list for since how would you be able to know which chapter it was thrown into?
At the end of the day I feel like a cookbook should be an easy tool for someone to use in the kitchen when there’s already so much going on. The book is visually exhausting to read, but looks very very delicious to make from.
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Reviewed in the United States on October 11, 2023
★★★★★ 3
Nice recipes, AWFUL font
Format: Hardcover, Format: Hardcover
I loved Cook This Book. Mine has a permanent crease on the Caesar salad page and the other recipes I have tried have all been winners, so I was excited to get Molly’s next book. My initial browse through the recipes has me excited to try them out, I just cannot understand the choice of this awful font. It is so difficult to read and make out the letters. I can’t imagine how difficult it would be for someone with visual or reading impairments. There is even a fully black page with dark blue font that is basically unreadable (the picture I included is very kind, in real life the contrast is extremely minimal). It’s such a huge distraction from what otherwise appears to be a nice collection.
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Reviewed in the United States on October 12, 2023
★★★★★ 5
Useful
Format: Kindle
Wonderfully written with lots of good advice. I think a lot of young people are lost in the world because a lot of this stuff is not taught in school anymore, at least not where I am. This would be a great resource for kids that are aging out of foster care and need the guidance of how to manage life. This could also serve to guide adults leaving prison.
I wish I would have had something like this when I was becoming an adult because I didn't have the best guidance and figured a lot of stuff out on my own. I will make sure both my kids have a copy to guide them in the event I'm not around to help them. Overall, great book and I'm glad someone wrote it. Lol
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Reviewed in the United States on November 26, 2024