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AB277717

Human C4b ELISA Kit (Complement C4-B)

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Human C4b ELISA Kit (Complement C4-B) is a single-wash 90-min Simplestep used to quantify Human C4b (Complement C4-B) with a sensitivity of 34.83 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Validated on a number of sample types including cerebrospinal fluid (CSF)
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

CO4, CPAMD3, C4B_2, C4B, Complement C4-B, Basic complement C4, C3 and PZP-like alpha-2-macroglobulin domain-containing protein 3

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ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)
  • ELISA

Supplier Data

ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)
Sandwich ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)
  • sELISA

Supplier Data

Sandwich ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)

Serum diluted 1 : 100,000 from ten individual healthy human female donors was measured in duplicate.

Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean C4b concentration was determined to be 299.35 μg/mL with a range of 95.86 – 529.04 μg/mL.

Sandwich ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)
  • sELISA

Supplier Data

Sandwich ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)

Interpolated concentrations.

Undiluted samples are as follows : saliva 1 : 25, HepG2 supernatant 1 : 20, urine 1 : 50, cerebrospinal fluid 1 : 500, and milk 1 : 500. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean C4b concentration was determined to be 135.80 ng/mL in saliva, 103.10 ng/mL in HepG2 supernatant, 408.43 ng/mL in urine, 2319.81 ng/mL in cerebrospinal fluid, and 3468.94 ng/mL in milk.

Sandwich ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)
  • sELISA

Supplier Data

Sandwich ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)

Example of human C4b standard curve in Sample Diluent NS. The C4b standard curve was prepared as described in Section 10.

Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)
  • sELISA

Supplier Data

Sandwich ELISA - Human C4b ELISA Kit (Complement C4-B) (AB277717)

Interpolated concentrations of native C4b in human serum and plasma samples.

The concentrations of C4b were measured in duplicates, interpolated from the C4b standard curves and corrected for sample dilution. Undiluted samples are as follows : serum 1 : 50,000, plasma (citrate) 1 : 25,000, plasma (EDTA) 1 : 50,000, and plasma (heparin) 1 : 50,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean C4b concentration was determined to be 351.14 μg/mL in serum, 52.76 μg/mL in plasma (citrate) and 307.91 μg/mL in plasma (EDTA), and 156.35 μg/mL in plasma (heparin).

Key facts

Detection method

Colorimetric

Sample types

Cerebral Spinal Fluid, Saliva, Urine, Heparin Plasma, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 34.83 pg/mL

Range

140.63 - 9000 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human C4b ELISA Kit (Complement C4-B) ab277717 is a rapid single-wash 90-min Sandwich ELISA to measure Human C4b (Complement C4-B)in cell culture supernatant, CSF, citrate plasma, EDTA plasma, heparin plasma, milk, saliva, serum, urine. This SimpleStep sensitivity is 34.83 pg/mL.

How the assay works

Human C4b (Complement C4-B)SimpleStep ELISA®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.

Assay Specificity

Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human C4b ELISA Kit (Complement C4-B) ab277717 protocol summary

1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm

C4b is a non-enzymatic component of the C3 and C5 convertases and essential for the propagation of the classical complement pathway. C4b covalently binds to immunoglobulins and immune complexes and enhances the solubilization of immune aggregates and the clearance of IC through CR1 on erythrocytes. C4b catalyzes the transacylation of the thioester carbonyl group to form ester bonds with carbohydrate antigens. Mouse and rat share 76 % and 79 % sequence homology with human C4b.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Serum", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "3.9" }, { "reproducibilityType": "Intra", "sample": "Serum", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "1.9" } ]
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Recovery

[ { "sample": "Cell culture supernatant", "range": "94 - 118 %", "average": "= 107" }, { "sample": "Milk", "range": "91 - 94 %", "average": "= 92" }, { "sample": "Serum", "range": "89 - 95 %", "average": "= 92" }, { "sample": "EDTA Plasma", "range": "92 - 96 %", "average": "= 93" }, { "sample": "Cerebral Spinal Fluid", "range": "102 - 109 %", "average": "= 106" }, { "sample": "Urine", "range": "99 - 118 %", "average": "= 109" }, { "sample": "Saliva", "range": "80 - 89 %", "average": "= 88" }, { "sample": "Heparin Plasma", "range": "93 - 100 %", "average": "= 96" }, { "sample": "Citrate plasma", "range": "97 - 103 %", "average": "= 101" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The 'C4b' protein also known as the 'C4b complement' plays a significant role in the immune system. This protein is a part of the larger complement system specifically as a segment derived from the cleavage of the native 'C4 complement'. It has an estimated molecular mass of about 200 kDa. The 'C4b' protein is prominently expressed on the surface of cells and in body fluids where the immune response is active. As an important player in the complement pathway it binds covalently to foreign particles labeling them for destruction or removal by immune cells.
Biological function summary

The 'C4b complement' interacts with various other components of the immune system. It is particularly involved in the formation of the C3 and C5 convertase complexes which are essential for the opsonization and activation cascade leading to pathogen elimination. Its attachment to cellular surfaces provides a docking point for proteins such as C2 facilitating the further cleavage and progression of the complement cascade. This interaction is critical for driving the classical and lectin pathways of complement activation.

Pathways

The 'C4b protein' is integral to the classical pathway of complement activation as well as to the lectin pathway. It associates with proteins like C1s C1q and factor B forming complexes that execute various stages of immune defenses. By forming C3 and C5 convertase 'C4b' not only promotes phagocytosis but also inflammation and cell lysis through membrane attack complexes. These processes underlie the body's ability to address microbial invasions efficiently.

'C4b complement dysregulation' can lead to autoimmune conditions such as systemic lupus erythematosus (SLE) and type 1 diabetes. In SLE reduced or malfunctional 'C4b' increases susceptibility to infections and triggers inappropriate immune responses. It can also correlate with the deficiency of related proteins like C2 which further impairs the complement system. These associations indicate that proper regulation of 'C4b' is essential for maintaining immune system stability and preventing overactive immune responses.
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Product protocols

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Target data

Non-enzymatic component of the 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.
See full target information C4B
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websiteProtocolBooklet
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