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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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4.0 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
Awesome Shelf for a Great Price!
The shelf is very sturdy and built well. I did not expect the quality to be so good. I honestly don't think you could build one with parts from the hardware store for less money. It was not very difficult to assemble, but it would have been easier if the were unions used with the pipes. I had to turn the whole shelf to tighten it. I also added some lights and the wires were difficult to snake through the pipes but I had planned on that not being easy. I did need help hanging the shelf. I don't think it can be done without help. I am extremely happy with the product and have got several compliments. Now I have to buy one for my mom because she likes mine so much. Shipping was super fast, it got to my house 1 day before I expected it. Thanks, I will be buying another one as soon as it gets back in stock
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Reviewed in the United States on November 2, 2020
★★★★★ 4
Looks great, not super sturdy
Size: 6 tier, Color: Black Brush Silver, Size: 6 tier, Color: Black Brush Silver
Putting the shelf together was simple and it looks nice. The shelf is not super study though. Word of advice, DO NOT use the drywall anchors that come with the shelf; buy your own. The instructions say to use a 5/16 drill bit and the included anchors. The anchors are terrible; the 5/16 bit was too big and the anchors went right through the wall when we tried to put the screws in. It was a huge frustration to put eight holes in the wall that we then had to patch up and repaint over because the included materials are terrible. Luckily a couple of the screws line up with the studs so it's fairly sturdy, but use our own anchors.
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Reviewed in the United States on January 25, 2022
★★★★★ 5
So good I bought two
Size: 6 tier, Color: Black Brush Silver
These shelves are GREAT. Why? They arrive in a compact well protected package, they go up in minutes, are super tall, and they look fantastic. The price is perfect. The components are not 'real' iron pipes (I've built my own iron pipes shelves before and it's extremely heavy), but some kind of lighter weight metal. That's fine by me! The shelves appear to be real wood (!!). If you buy cheap furniture you know what fake wood looks like. You can tell. Guests can tell. This is probably a pretty cheap wood but who cares--it's REAL, is stained nicely with a distressed look, and overall looks amazing once books and items go on it.
If you are deciding between these and some cheaper similar thing, pay a biiiit more and get something with real wood. I just bought a second for my office and they'll hold my books, art, and speakers.
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Reviewed in the United States on May 28, 2021
★★★★★ 5
Perfect. Sturdy. Super easy to assemble.
Size: 6 tier, Color: Black Brush Silver
I don’t rate a lot of items but I’m in love with this one. I just received it today. It was packaged incredibly well, took 15 min or less to put together and looks amazing!!! I’m tempted to order another one for another room in the house. It’s sturdy, well-made and I can’t stress how easy it was to assemble. Great product, highly recommend. (The owners even sent a hand-written note thanking me for ordering - what a nice touch!)
Edit: ordered a second one after living first one so much
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Reviewed in the United States on December 30, 2020
★★★★★ 5
Perfect industrial look bookshelf
Size: 6 tier, Color: Black Brush Silver, Size: 6 tier, Color: Black Brush Silver
Surprised how good the quality was for the price. Metal is real metal and wood seems to be real wood. Pretty much twist on and off each section to put together. No question it is a very sturdy piece of furniture. Bought it over other bookshelves because the reviews said it was solid … and indeed it did turn out to be very solid. I’m very pleased and would recommend. Now I just need some things to fill the shelves.
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Reviewed in the United States on May 18, 2022