AB277717

Human C4b ELISA Kit (Complement C4-B)

Recombinant
SimpleStep
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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

別名を表示する

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

5 Images

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

ELISA

Supplier Data

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.

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.

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.

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

検出方法

Colorimetric

サンプルタイプ

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

交差種

Human

アッセイタイプ

Sandwich

Quantitative

検出感度

= 34.83 pg/mL

検出範囲

140.63 - 9000 pg/mL

アッセイ時間

1h 30m

アッセイプラットフォーム

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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製品の詳細

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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精度

[ { "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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回収率

[ { "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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製品内容

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出荷温度及び保存条件

出荷温度

Blue Ice

短期保存温度

+4°C

長期保存温度

+4°C

保管に関する情報

+4°C

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補足情報

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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製品プロトコール

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ターゲットの情報

Precursor of non-enzymatic components of the classical, lectin and GZMK complement pathways, which consist in a cascade of proteins that leads to phagocytosis and breakdown of pathogens and signaling that strengthens the adaptive immune system.. Complement C4b-B. Non-enzymatic component of C3 and C5 convertases (By similarity). Generated following cleavage by complement proteases (C1S, MASP2 or GZMK, depending on the complement pathway), it covalently attaches to the surface of pathogens, where it acts as an opsonin that marks the surface of antigens for removal (By similarity). It then recruits the serine protease complement C2b to form the C3 and C5 convertases, which cleave and activate C3 and C5, respectively, the next components of the complement pathways (PubMed : 8538770). Complement C4b-B isotype catalyzes the transacylation of the thioester carbonyl group to form ester bonds with carbohydrate antigens, while C4b-A isotype is responsible for effective binding to form amide bonds with immune aggregates or protein antigens (PubMed : 8538770).. C4a anaphylatoxin. Putative humoral mediator released following cleavage by complement proteases (C1S, MASP2 or GZMK, depending on the complement pathway). While it is strongly similar to anaphylatoxins, its role is unclear. Was reported to act as a mediator of local inflammatory process; however these effects were probably due to contamination with C3a and/C5a anaphylatoxins in biological assays.

See full target information C4B

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websiteProtocolBooklet

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Human C4b ELISA Kit (Complement C4-B)