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Description
Human CFHR1 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 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. 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 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 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 7 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 directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 capture antibody against Complement Factor H Related Protein 1 (CFHR1). After incubation and washing, the color is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Complement Factor H Related Protein 1 (CFHR1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Complement Factor H Related Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Complement factor H-related protein 1 is encoded by the CFHR1 gene. It binds to the Pseudomonas aeruginosa elongation factor Tuf and is activated by the protease. Tuf functions as a virulence factor by acquiring host proteins onto the pathogen surface, controlling complement, and promoting tissue invasion. Mutations in this gene are associated with an increased risk of atypical hemolytic uremic syndrome. Diseases associated with CFHR1 include hemolytic uremic syndrome, atypical 1, and age-related macular degeneration 1. An important homologue of this gene is CFHR2. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, and other biological fluids |
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4.4 ★★★★★
Based on 14 reviews
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Product Reviews
★★★★★ 4
Kiss the Fae
Format: Kindle
I have to be honest, I have spent the last 24 hours going back and forth between three and four stars. Ultimately, I enjoyed the story and I think that is all that counts. So, 4 stars it is!
What I loved, Labyrinth is my favorite movie of ALL TIME! Growing up I dreamed about marrying David Bowie and thought Jareth was the sexiest man of all time. So, I was tickled to read a story loosely based on the movie. And it did not disappoint! Natalia Jaster did such a good job following along with the story while making it completely her own.
I try not to give spoilers in my reviews—but—let’s just say I loved Cerulean, BUT I AM SOOOOO EXCITED ABOUT HIS BROTHERS STORIES!
Now the not so popular opinions . . .
What bugged the ever lovin’ crap out of me . . . and I say this with all the love and respect in the world for the author and all the hard work that went into this book—but girl!!! There were SO many times where unnecessary synonyms were used to the point it would almost make the sentence not make sense. I wanted to shake my kindle and ask to be a Beta reader so I could help avoid this. It does not make you look like a bad writer to use simple words when simple words feel right to the flow of the story. End of tangent.
The other thing that was not my jam was the verb tense. I am not a huge fan of present tense, but that is on me and did not take away from my rating or the enjoyment of the story. Just a thing that would randomly catch me off guard and pull me out of the story.
Lastly, there are a few times (especially in the beginning) where our MC Lark is awkwardly slutty; to the point it feels forced. You can have a promiscuous character without making her talk about sex randomly in the middle of a conversation. But be warned, this book brings the smut! It is New Adult/Adult NOT YA!
Overall, I enjoyed it and very much look forward to the next book that comes out in April!
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Reviewed in the United States on February 10, 2021
★★★★★ 5
Wonderful Wicked Fantasy
Format: Kindle
Kiss The Fae (Dark Fables :Vicious Faeries Book 1) - Natalia Jaster
I originally read this book before the edited version bumping this to a more yummy book with Adult content and it was well worth the wait.
Nadia Jaster has become one of my favorite authors and I have read all her books some of them twice.
The beautiful descriptions and almost poetic world and character building drew me in and immersed me in this fantasy realm.
Lark is a female who I adore, actually all three “sisters” are amazing in their own way. Lark is adventurous , a free spirit who embraces her sexuality and freedom like a crown. Lark is summoned to Faerie and ends up in the court of the sexy and pretty (her favorite kind) Cerulean, Lord of the Sky (and whom she had met and rescued 9 years ago when they were both youngsters)
Cerulean rules the wind and sky and must have Lark solve a difficult task to gain her freedom. It is hard not to fall in love with the beautiful Fae who speaks in wicked riddles that are both enticing as well as seducing.
The two are a wonderful pair whose slow burn love builds enticingly throughout the book. Once those flames are built their relationship is magical and guaranteed to melt many panties. Their love and heated scenes are written beautifully. Many circumstances pulled at my heartstrings as well as humor, excitement, danger, loyalty, adventure and romance with a big dreamy “Sigh”.
The lore of the Faeries is traditional in many details as well as original and unique storylines.
The first book of the series is a phenomenal and enchanting start you will not want to leave.
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Reviewed in the United States on July 19, 2022
★★★★★ 5
Fabulously Sexy with Serious Story Depth
Format: Kindle
Cerulean is incredibly yummy, and Jaster writes sex like nobody’s business. But the real highlight of the tale is what she always manages to do: make the story actually mean something.
There are themes revolving around animal rights, how devastating animal cruelty is, racial division and derision, how important change can be in a society that thinks it has it all figured out, and how the smallest pebble tossed into the pond can create lasting ripple effects that change everything.
I love this author, I love her poetry of imagination, I love her sassy, witty characters, her dark heroes, and her ability to write characters that recognize when they’ve been wrong, even if it takes them a while to figure it out.
This book was captivating to the last word. Completely enchanting, as a book about the Fae only ever should be.
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Reviewed in the United States on November 12, 2020
★★★★★ 4
i want to give this a 5 star BUT...
Format: Paperback
So, it took almost a month to get this series delivered, which is fine. I was just super excited to start it. But right when I got them and started to read the first book (KISS THE FAE) the first page fell out, then the 3rd page, and I had to tape the binding on either end just so no more pages fell out...that being said. THE BOOK IS PHENOMENAL!!!!! The story line *chefs kiss* the details and the overall descriptive nature of her writing style *chefs kiss*! I am in love with the story (so far) and after 2 days I am near the end of book 1. I am highly anticipating the remainder of this series. But I just wish I could give it 5 stars (for the mere fact the pages were falling out, I gave it a 4 star) BUT DEFINITELY READ THIS SERIES - SOOOOOOOOOOO GOOD!!!!
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Reviewed in the United States on January 23, 2024
★★★★★ 5
Fallen Angels, fae, vampires, oh my!
Format: Kindle
Rating: 4.5 | Spice: 2 (but a good slow-burn)
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Main Characters: Huntyr and Wolf
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I couldn’t wait to read this book; there was so much hype about it! And there was no doubt why. I fell in love with the characters and the plot itself. This book is mainly plot driven more than friction driven but it’s easy to follow along with. The characters are fun, easily understood. The main setting is at an academy where both the main characters are going through trials and building strength for the final test, The Transcendent. There are fantastic side characters as well. I loved the camaraderie between Huntyr and her friends. But we don’t like Lanson. 😆
We do have some plot twists that come into play throughout the book. Secrets and betrayal to be seen. I did adore Wolf and Huntyr’s relationship. It was a classic slow burn trope. They didn’t hit it off fast, but in time their feelings grew. I loved their banter, so sexy. Wolf is your next book boyfriend; Huntyr is your next vampire assassin independent bad-a*s female. Themes include loyalty, trust, self-discovery, a true slow burn romance. Side note: book ends on a angsty cliffhanger!
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Emily, thank you for writing this awesome novel and I cannot wait to devour Book 2, Blood So Brutal! 😍
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Happy reading, my lovelies! xo
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Reviewed in the United States on January 21, 2024