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Description
Rat FX ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge at 1000g for 15 minutes at 2 8C within 30 minutes of collection. Remove the supernatant for testing or store at
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. 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. 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 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 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 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 Coagulation Factor X (FX) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Coagulation Factor X (FX) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Coagulation Factor X ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Coagulation factor X (FX), also known as Stuart-Prower factor, is an enzyme in the coagulation cascade. It is a serine endopeptidase (proteinase group S1, PA family). Factor X is synthesized in the liver and requires vitamin K. FX is activated to factor Xa through hydrolysis by factor IX (in complex with its cofactor factor VIII, known as the intrinsic tenase) and factor VII (in complex with its cofactor tissue factor, known as the extrinsic tenase). | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | plasma |
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4.5 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 3
Solid 3.5 for me.
Format: Kindle
The story in the characters were unique which I liked. Never read a story like it. Kept me interested. It just wasn't as captivating as I hoped.
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Reviewed in the United States on May 25, 2026
★★★★★ 4
🖤🖤4 Stars🖤🖤
Format: Kindle
“There are darknesses in life and there are lights. You are one of the lights...The light of all lights.” - Master of Salt & Bones.
REVIEW: This book follows Isa who lives with her aunt on an island. Isa begins a new job as a caretaker for the dark, and mysterious Blackthorne family. A family who is disliked and even feared by the rest of the inhabitants of the island. However, as she grows closer to the man who they call the devil, Lucian Blackthorne, Isa sees for herself what the truth of Blackthorne manor really is.
This book is a dark romance! Originally I was told it was fantasy, which I don’t agree with at all. It is more of an erotic, gothic feel with some mystery sprinkled in. The plot was very interesting and kept me intrigued. This book was also quite steamy, however, I felt those scenes were well done and didn’t take away from the rest of the book.
Overall, the content in this book is quite dark and the backstories of the characters involve potentially triggering matters. Be aware to look these up if you feel like you need to. Overall, this is the first book I have read by Keri Lake, and I am excited to read more.
SYNOPSIS: When I was a little girl, I dreamed a handsome knight would come and rescue me from my wretched mother. He'd ride up on his white steed and break the curse I've been fated to carry since the day I was born.
Funny how things changed over time. How the fairy tale twisted into something far more crooked, darker than I ever imagined.
In reality, my knight is scarred and broken, living alone in a castle of bones that overlooks the sea. He isn’t searching for me. He never was.
Lucian Blackthorne is as cursed as I am, and equally shunned by the locals, the fishers of men, who believe him to be the devil in the flesh. Perhaps he is, with the way his amber eyes draw me in, ignite me like an infernal blaze. And the sins he whispers in my ear are as wickedly intoxicating as the man himself.
Yet, his touch is heaven and his will is my weakness. He calls us forbidden, an unsalvageable tragedy, with no happy end. Maybe we are. But in this story, he’s the one who needs saving.
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Reviewed in the United States on August 11, 2020
★★★★★ 5
one of my top reads of 2023 🖤
Format: Paperback, Format: Paperback
Master of Salt & Bones by Keri Lake review (no spoilers)
♾️/5 ⭐️
4/5🌶️
🏰gothic dark romance
🎹murder/mysteries
🏰age gap
🎹spicy
🏰touch her & d!e
🎹secret society
🏰dual POV & a past timeline for Lucian
“we’re the composers, the conductor’s of our own fate, and we write the notes to a beautiful, dark melody that no one else can hear.”
this book was AMAZING ❤️🔥
i couldn’t put this book down for anything .
this book gives beauty & the beast / phantom of the opera vibes with a lot more going on . Lucian hires Isadora to help take care of his mom, not his nurse but a companion for her . Isadora has a painful past & she’s willing to do anything to get out of town to make some money , even if that means working for a guy with a thousand bad rumors about him . the way Lucian talks to Isadora is like he’s speaking poetry without even trying . the wording & quotes in this book were chiefs kiss . the spice in this book was gold 🔥 the plot of this book is super original & sooooo twisty . i think there were 5 different twists at the end & i had my jaw on the floor at all of them .
literally obsessed with this book . the writing was beautifully done 🫶🏼
this is my first book by Keri & i will now be ordering more from her immediately🖤
this book is definitely in my top reads of the year!
as always check your TW 🤍
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Reviewed in the United States on September 29, 2023
★★★★★ 5
So Worth the Read
Format: Audiobook
I absolutely loved this book. The pacing was great, the characters felt real, and the story kept me hooked the entire time.
There were a few moments that genuinely surprised me, and I appreciated how everything came together at the end. Highly recommend for anyone who enjoys Dark Romance/ Gothic Fiction
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Reviewed in the United States on March 4, 2026
★★★★★ 4
4 star read - dark gothic romance
Format: Kindle
Master of Salt and Bones by Keri Lake
4 STAR READ
“You’re my curse. Staying away from you, is like trying to hold my breath when the tide is rising.”
This book reminds me a lot of Nightshade, also by Keri Lake, but without the fantasy element. It gives off the same gothic vibes, including flash backs, history of abuse, and sexual deviances. It also has some overlap with the older male character obsession that is seen in Nocticadia - though Lucien doesn’t hold a candle to Professor Bramwell.
SUMMARY
Lucien was abused, lied to, and manipulated his whole life, by everyone who should have protected and cared for him. A wealthy heir to a massive fortune, he is hard, dark, and delicious. The story unfolds across two timelines, jumping between Lucien’s past and present, and Isa’s POV. Lucien is viewed as the devil of Tempest Cove, covered in scars and rumors abound involving the death of his wife and son. Isa is the daughter of the town whore, who has struggle to fit in and cope with the tribulations of her life. Lucien hires Isa to be his mother’s companion, to keep her company and encourage her to get out of bed each day. As they continue to bump into each other and interact, their relationship develops beyond his dark obsession. Lucien is wary to get attached as his connection to a dangerous secret society puts everyone he cares about at risk, however there may be no way around Isa’s involvement.
WHAT I LIKED and DISLIKED
It was a great story full of tragedy and resilience. Both characters have deep back stories that unfold throughout the book, we get information at just the right time to make things make sense. It is really well told. It does feel a bit slow in the middle, I took a mid book break to read something else but came back to it and wondered why I stopped - the reason I gave 4 stars was because I had the need to take the break. Which tells me I lost interest during part of it.
Lucien has so many layers and each one is full of sadness and devastation. Isa’s story is tragic in its own way, however her youth has her dealing with them in a difference way - she is definitely more detached in her own way.
There is an age gap - Lucien is 33, Isa is 19 — I’m not usually a fan of this kind of age gap, but Isa isn’t childish or immature so it works. The book definitely gives Grumpy x sunshine vibes. The spice is well done and paced well throughout the book.
If you like spicy, gothic, dark romance and have enjoyed other books by Keri Lake this book will not disappoint.
There are references to rape, sexual abuse, physical abuse, and mental illness — I would check any triggers before reading
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Reviewed in the United States on June 17, 2025
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