SKU: 55746085094

Human C3a ELISA Kit

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Description

Human C3a 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 the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing.
3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed.
4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis.
5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles.
6. 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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.
Sensitivity 0.053 ng/mL
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 Complement Component 3a (C3a) 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 Complement Component 3a (C3a) 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 Component 3a  ELISA Kit
Detection Type Double antibody sandwich method
Composition
Name 9 6 T  match   set remark
Pre-coating 96 Well plate 8 Hole ×12 Strip without
Standard 2 branch
Dilute as per instructions
Universal diluent
2×20mL
without
Concentrated biotinylated antibody ( 100× )  
120uL
Dilute as per instructions
Concentrated enzyme conjugate ( 100× )
120uL
Dilute as per instructions
20× Washing liquid
2×10mL
Dilute as per instructions
Bottom thing ( TMB )
10mL
without
Stop liquid
6mL
without
Sealing film
4 Zhang
without
Instructions
1 Share
without
Background Complement fragment 3a (C3a) is one of the proteins formed by cleavage of complement component 3; the other is C3b. C3a is a passive 77-residue toxin that binds to the C3a receptor (C3aR), a class A G protein-coupled receptor. It plays a major role in immune responses. C3a molecules induce responses through the GPCR C3a receptor. Like other aprotinins, C3a is regulated by cleavage of the arginine at its carboxyl terminus, generating a molecule with less inflammatory activity (C3a desarg). It is an effector of the complement system with a range of functions, including T cell activation and survival, stimulation of angiogenesis, chemotaxis, mast cell degranulation, and macrophage activation. It has been shown to have both pro-inflammatory and anti-inflammatory effects, with its activity counteracting the pro-inflammatory effects of C5a.
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.
Storage Temp. If the unopened kit is stored at 4°C, the shelf life is 6 months.
Test Range 0.156-10 ng/mL
Applications Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids
Shipping Notes
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Exchange/Return Notes
  • We offer a 30-day return/exchange service after receiving.
  • Final sale items are not eligible for returns or exchanges.
  • To process your return/exchange, please contact us at [email protected]
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SKU: 55746085094

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Mark
West Palm Beach, US
★★★★★ 5
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Size: 8.5-9.5 Women/7.5-8.5 Men, Color: Shark Blue
Perfect! I’m autistic and this was the last piece of the puzzle for my sensory experience at the pool. They feel like wearing socks with hard soles on and are great for if you need to walk through gravel or anything. They don’t trap water much and the fabric is stretchy and comfortable. I liked the range of colors, I got the one with sharks. I wear a men’s size 8 shoe and I got the same size here, it was the correct size with a little extra room.
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Reviewed in the United States on May 27, 2026
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Amazon Customer
Alexandria, US
★★★★★ 4
Colorful, comfortable, durable water shoes
Size: 8.5-9.5 Women/7.5-8.5 Men, Color: Shark Blue, Size: 8.5-9.5 Women/7.5-8.5 Men, Color: Shark Blue
I have ordered 5 pairs of these water shoes. The quality is amazing the bottom is durable, the stitching feels strong, they are comfortable, slip resistant, fit true to size and look awesome! We ordered the paisley, the blue geo, galaxy, lightning (lightning was sent back as we received the wrong size) and sharks. Only reason they are getting a 4 star instead of 5 is because I would much rather have the cushion inside sewn in instead of it being like an insert but that is what I find to be most common with these type of shoes.
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Reviewed in the United States on April 29, 2026
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Susan L.
Omaha, US
★★★★★ 5
Love these for beach foot protection!!
Size: 10-11 Women/9-10 Men, Color: Black-w001
These water shoes were perfect for our cruise to Grand Turk. The beach was a little rocky once in the water & the bottom of these shoes protected our feet and made it comfortable they dry quickly they are lightweight and easy to pack and carry in a beach bag. I bought two pair of the black one for me one for my husband, and he even wore them which was shocking into me and loved them.!!!! they don’t have walking support so they’re definitely something that you bring with you put on your feet before you enter like water where you can’t see the bottom or rocks/coral and they totally protect your foot on the bottom
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Reviewed in the United States on May 13, 2026
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TB
Belleville, US
★★★★★ 3
I received someone else’s returned product
Size: 7-8 Women/6-7 Men, Color: Sunset
The inner product packaging was already cut open and not sealed, and there was debris inside the socks: clear indicators that the pair I received was someone else’s return. They also run big and are very wide in the toe area. My daughter’s pair also ran big, but at least hers were sealed. Overall quality is a bit flimsy, so we’ll see how long they last.
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Reviewed in the United States on May 31, 2026
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Waukegan, US
★★★★★ 5
Decent water shoe
Size: 10-11 Women/9-10 Men, Color: N-laser Stripe
I have to admit, I was a bit skeptical about the quality of and comfort of these shoes because of the cheap price. However, I really do like them. They stay on my feet, are comfortably and non-binding, which is saying something because my feet have been pounded over the years. They even did well when I had to cross some rocks, trekking back to the kayak. Definitely a gods water shoe for a good price.
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Reviewed in the United States on May 25, 2026

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