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Description
Mouse C5aR1 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 preparation and requirements: Tissue homogenization: Rinse the tissue with pre chilled PBS (0. 01M, pH 7. 4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and
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 preparation and requirements: Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 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 Complement Component 5a Receptor 1 (C5aR1). After incubation and washing, the assay 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 Complement Component 5a Receptor 1 (C5aR1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Complement Component 5a Receptor 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The C5a receptor, also known as complement component 5a receptor 1 (C5AR1) or CD88 (cluster of differentiation 88), is a G protein-coupled C5a receptor. It functions as a complement receptor. The C5a receptor regulates inflammatory responses, obesity, development, and cancer. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.9 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Love it
Size: 11, Color: Black
Almost the perfect travel shoe. Goes well with both casual and formal attire. Walked like a million steps with this in Japan, SEA, etc. only thing it probably can't do is probably hiking.
Otherwise the leather feels really high quality, and the shoe slips on without much fuss. I'd describe the style as: a dressed-up Converse
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Reviewed in the United States on March 22, 2026
★★★★★ 1
Would not every buy marc Joseph shoes ever
Size: 11 Wide, Color: Black
The marc Joseph shoes did not hold up had a blow out rightside of shoes had the shoes for about two weeks when my wife noticed and say that not good.i buy Cohen shoes and Clarke shoes
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Reviewed in the United States on May 7, 2026
★★★★★ 2
Great shoe, horrible lace design.
Size: 13, Color: White, Size: 13, Color: White
This shoes look great and the materials for the actual shoe feel great. They run a little big that doesn't bother me. Unfortunately, the lace design is horrible. I've now have 2 pairs of these shoes and both times the bottom lace has ripped out of the stitching. The first time was on the 2nd day I wore them, and the replacement pair to those it happened after I walked 20 feet. Both times I've tied a knot in the bottom lace so that I could get through the day. Ideally these laces should be connected together and not attached to the shoe (this is how both of my Skechers Slip-ins are designed). Because I like the rest of the shoe so much I'm buying elastic laces to replace the horrible original laces.
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Reviewed in the United States on March 14, 2025
★★★★★ 5
The "Lazy Professional" Look: Is Hands-Free Luxury Actually Real?
Size: 9.5, Color: Black Napa Leather
Living down here in Florida, my footwear needs are pretty specific. It’s hot, it’s humid, and I spent half my life rushing from the car into work or a meeting. I’ve reached that age where I value efficiency just as much as style—maybe more. I’ve been eyeing the Marc Joseph New York Hands-Free Slip-on Penny Loafers for a while, and after putting them through the wringer, here is the honest truth from someone who just wants to look sharp without the hassle.
The "Just Step-In" Reality
Look, the big selling point here is the "Hands-Free" tech. We’ve all seen the commercials for those athletic slip-ins, but finding that in a legitimate leather penny loafer is a different game. Does it work? Yes, surprisingly well. The heel counter is firm enough that it doesn’t collapse when you slide your foot in, but it doesn’t feel like a piece of plastic digging into your Achilles once you’re in. For those of us who are tired of bending over or hunting for a shoehorn every morning, this is a genuine quality-of-life upgrade.
The Florida Factor: Comfort and Style
The leather is actual calfskin (on most models), which is a must for the Florida heat. Synthetic shoes turn into a sauna within ten minutes, but these breathe reasonably well. The aesthetic is classic—it’s a "professor" shoe through and through. You can wear them with chinos and a blazer for work or throw them on with some nice jeans for a weekend lunch.
Inside, they’ve got a gel heel insert and a padded footbed. It’s not quite "walking on a cloud"—let’s not over-hype it—but it’s a massive step up from the hard, flat soles of traditional dress loafers. I’ve spent four hours on my feet lecturing, and my arches didn’t hate me by the end of the day.
The Sizing Gamble
Here’s where you need to be careful. The consensus from other guys (and my own experience) is that the sizing is a bit of a coin toss. They tend to run a little large and sometimes wide. If you have narrow feet, you might find the sides "gaping" or flaring out when you walk, which kills the sleek look. I’d recommend ordering a half-size down if you’re usually between sizes.
The Breakdown
The Pros:
True Hands-Free: You can actually put these on while holding a coffee and a briefcase. No hands needed.
Legit Materials: The calf leather feels premium and smells like the real deal.
Versatility: Perfectly bridges the gap between a "car shoe" and a formal loafer.
Comfort: The gel insert makes a noticeable difference for long-duration wear.
The Cons:
Sizing Inconsistency: They can run wide, leading to some "heel slip" if you don't get the perfect fit.
Break-in Period: The heel is stiff by design (to allow for the slip-on feature), so expect a day or two of minor stiffness.
Price Point: They aren't cheap, though often on sale.
Value for Money
Are they worth it? If you’re comparing them to high-end Italian brands that cost $500, these are an absolute steal. They look 90% as good for a fraction of the price. However, if you're used to $60 mall shoes, the jump to $150+ might feel steep until you realize you’re paying for the convenience of never having to touch your shoes to put them on. For a daily driver in a professional setting, the value is definitely there.
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Reviewed in the United States on April 10, 2026
★★★★★ 5
Step In Look Good
Size: 11.5, Color: Cognac Napa Leather
I am wearing these shoes as I write this review, and they look and feel great. I have a bit of a disability with drop foot on the right leg and bilateral neuropathy and have been wearing step in shoes for about 10 years. But the first company that introduced leather dress and casual shoes stopped making them and now only makes sport and casual shoes. So I have been searching for some that meet my need for shoes like this and this company has given me three pairs, so far. Easy to get into, comfortable to wear, and good looking. And they take polish very well, too. (Remember how to do that?). We may buy another pair or two in different styles as well.
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Reviewed in the United States on April 26, 2026