Human C4a ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human C4a in Serum, EDTA Plasma, Citrate plasma, Heparin Plasma, Milk samples.
Application | Reactivity | Dilution info | Notes |
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Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
Non-enzymatic component of C3 and C5 convertases and thus essential for the propagation of the classical complement pathway. Covalently binds to immunoglobulins and immune complexes and enhances the solubilization of immune aggregates and the clearance of IC through CR1 on erythrocytes. C4A isotype is responsible for effective binding to form amide bonds with immune aggregates or protein antigens, while C4B isotype catalyzes the transacylation of the thioester carbonyl group to form ester bonds with carbohydrate antigens. Derived from proteolytic degradation of complement C4, C4a anaphylatoxin is a mediator of local inflammatory process. It induces the contraction of smooth muscle, increases vascular permeability and causes histamine release from mast cells and basophilic leukocytes.
CO4, CPAMD2, C4A, Complement C4-A, Acidic complement C4, C3 and PZP-like alpha-2-macroglobulin domain-containing protein 2
Human C4a ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human C4a in Serum, EDTA Plasma, Citrate plasma, Heparin Plasma, Milk samples.
Sample | n | mean | SD | C.V. |
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Sample Serum | n 8 | mean - | SD - | C.V. = 1.2 |
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Serum | n 3 | mean - | SD - | C.V. = 4.9 |
Sample type | Average % | Range |
---|---|---|
Sample type Serum | Average % = 100 | Range 90 - 108 % |
Sample type EDTA Plasma | Average % = 108 | Range 100 - 115 % |
Sample type Citrate plasma | Average % = 105 | Range 100 - 108 % |
Sample type Heparin Plasma | Average % = 92 | Range 85 - 103 % |
Sample type Milk | Average % = 104 | Range 101 - 106 % |
Human C4a SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of C4a protein in Serum, EDTA Plasma, Cit Plasma, Hep Plasma, and Milk. Quantitate Human C4a with 4.245 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab315065) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
Complement C4a also known as C4a anaphylatoxin is a component of the complement system with a molecular mass of approximately 9 kDa. This small protein originates from the proteolytic cleavage of complement component C4 which contributes to immune system activation. C4a is mainly expressed in the liver but can also be produced by other tissues during inflammation. It serves as a chemoattractant recruiting immune cells to sites of infection helping initiate a response against pathogens.
Complement C4a acts as an anaphylatoxin participating in the complex immune system response. Anaphylatoxins like C4a C3a and C5a contribute to the inflammatory process by inducing smooth muscle contraction increasing vascular permeability and triggering histamine release from mast cells and basophils. C4a complements the activities of other complement components playing an important role in modulating inflammation and defense mechanisms in the body.
The activation of C4a is an important step within the classical and lectin pathways of complement activation. These pathways involve the recognition and elimination of pathogens and unwanted cells. C4a along with its related proteins C4b and components C2 and C3 facilitates the formation of C3 convertase which amplifies the response and further clears pathogens. C4a also interacts with complement inhibitors which regulate these pathways to prevent excessive tissue damage during inflammation.
C4a anaphylatoxin has associations with autoimmune diseases and angioneoplasm. Changes in serum levels of C4a are observed in conditions like systemic lupus erythematosus where it may contribute to tissue damage. High C4a levels can correlate with disease activity and severity. Additionally C4a alongside other anaphylatoxins like C3a and C5a might play a role in angiogenesis influencing tumor progression and angiogenesis-related disorders serving as a potential biomarker for disease severity and prognosis.
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Example of human C4a standard curve. Background-subtracted data values (mean +/- SD) are graphed.
Interpolated concentration of native C4a was measured in duplicate at different sample concentrations. Undiluted samples are as follows: human plasma – heparin 1:1,600 and human milk 1:67. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Interpolated concentration of native C4a was measured in duplicate at different sample concentrations. Undiluted samples are as follows: human serum 1:10,000, human plasma – citrate 1:5,000, and human plasma – EDTA 1:6,400. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Serum of seven individual healthy human female donors was measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean C4a concentration was determined to be 4,579 ng/mL with a range of 991 – 7,804 ng/mL.
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