AB213472

Human IRAK4 ELISA Kit

Recombinant
SimpleStep
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Human IRAK4 ELISA Kit is a single-wash 90-min Simplestep used to quantify Human IRAK4 with a sensitivity of 247.4 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
- Different formats for different needs: 384-well plate for higher throughput
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

Interleukin-1 receptor-associated kinase 4, IRAK-4, Renal carcinoma antigen NY-REN-64, IRAK4

6 Images

ELISA - Human IRAK4 ELISA Kit (AB213472)

ELISA

Supplier Data

ELISA - Human IRAK4 ELISA Kit (AB213472)

Functional Studies - Human IRAK4 ELISA Kit (AB213472)

FuncS

Supplier Data

Functional Studies - Human IRAK4 ELISA Kit (AB213472)

Example of human IRAK4 standard curve.

Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human IRAK4 ELISA Kit (AB213472)

sELISA

Lab

Sandwich ELISA - Human IRAK4 ELISA Kit (AB213472)

Example of human IRAK4 standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.

sELISA

Supplier Data

Sandwich ELISA - Human IRAK4 ELISA Kit (AB213472)

Representative image of Human IRAK4 ELISA Kit (ab213472)

Components shown from left to right :

- 10X Wash Buffer PT (ab206977)

- 10X Human IRAK4 ELISA Kit Capture Antibody

- 10X Human IRAK4 Detector Antibody

- 10X Human IRAK4 ELISA Kit Lyophilised Purified Protein (2 vials)

- Human IRAK4 ELISA Kit 50X Cell Extraction Enhancer Solution (ab193971)

- Sample diluent NS

- TMB solution

- Stop Solution

- 5X Cell Extraction Buffer PTR (ab193970)

- Antibody Diluent 4BC

SimpleStep Pre-Coated 96-Well Microplate (ab206978) - shown behind bottles / vials.

Plate Seals - shown underneath bottles / vials.

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the ELISA kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit

sELISA

Lab

Sandwich ELISA - Human IRAK4 ELISA Kit (AB213472)

Interpolated concentration of native IRAK4 was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 125 µg/mL THP1 cell extract. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR.

sELISA

Supplier Data

Sandwich ELISA - Human IRAK4 ELISA Kit (AB213472)

Interpolated concentrations of native IRAK4 in human THP1 and PBMC extract based on a 500 μg/mL extract load.

The concentrations of IRAK4 were measured in duplicate and interpolated from the IRAK4 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IRAK4 concentration was determined to be 40.5 ng/mL in THP1 extract and 45.2 ng/mL in PBMC extract.

Key facts

Detection method

Colorimetric

Sample types

Cell culture extracts, Tissue Extracts

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 247.4 pg/mL

Range

0.5 - 32 ng/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (6 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 IRAK4 ELISA Kit ab213472 is a rapid single-wash 90-min Sandwich ELISA to measure Human IRAK4 in cell culture extracts, tissue extracts. This SimpleStep sensitivity is 247.4 pg/mL.

How the assay works

Human IRAK4 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 IRAK4 ELISA Kit ab213472 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

Save your precious samples

Our 384-well Human IRAK4 ELISA Kit only requires a maximum of 12.5µL sample

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-IRAK4 antibody [EPR19671-185] - BSA and Azide free (Capture) ab242833
- Anti-IRAK4 antibody [EPR19671-170] - BSA and Azide free (Detector) ab242833

IRAK4 is a serine/threonine protein kinase involved in the innate immune response. IRAK4 is recruited by MD88 through TLR signalling, where it phosphorylates IRAK1 and ultimately results in NF-kappa-B nuclear translocation and activation.The standard is affinity purified.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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Precision

[ { "reproducibilityType": "Inter", "sample": "THP-1 Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "4" }, { "reproducibilityType": "Intra", "sample": "THP-1 Extract", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "5" } ]

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Recovery

[ { "sample": "Cell culture extracts", "range": "107 - 115 %", "average": "= 111" }, { "sample": "Tissue Extracts", "range": "100 - 101 %", "average": "= 101" } ]

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What's included?

{ "values": { "1x384Tests": { "sellingSize": "1 x 384 Tests", "publicAssetCode":"ab213472-1x384Tests", "assetComponentDetails": [ { "size":"2 x 12 mL", "name":"Stop Solution", "number":"AB213472-CMP05" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB213472-CMP12" }, { "size":"2 x 1 Vial", "name":"Human IRAK4 Lyophilized Recombinant Protein", "number":"AB213472-CMP10" }, { "size":"1 x 600 µL", "name":"10X Human IRAK4 Capture Antibody", "number":"AB213472-CMP02" }, { "size":"1 x 6 mL", "name":"Antibody Diluent 4BC", "number":"AB213472-CMP09" }, { "size":"1 x 600 µL", "name":"10X Human IRAK4 Detector Antibody", "number":"AB213472-CMP03" }, { "size":"2 x 12 mL", "name":"TMB Development Solution", "number":"AB213472-CMP06" }, { "size":"1 x 1 mL", "name":"50X Cell Extraction Enhancer Solution (ab193971)", "number":"AB213472-CMP07" }, { "size":"1 x 1 Unit", "name":"Plate Seals", "number":"AB213472-CMP11" }, { "size":"1 x 50 mL", "name":"5X Cell Extraction Buffer PTR (ab193970)", "number":"AB213472-CMP08" }, { "size":"1 x 1 Unit", "name":"384 well CaptSure™ microplates", "number":"AB213472-CMP14" } ] }, "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab213472-1x96Tests", "assetComponentDetails": [ { "size":"1 x 12 mL", "name":"Stop Solution", "number":"AB213472-CMP05", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB213472-CMP12", "productcode":"" }, { "size":"1 x 1 mL", "name":"50X Cell Extraction Enhancer Solution (ab193971)", "number":"AB213472-CMP07", "productcode":"" }, { "size":"1 x 12 mL", "name":"Sample Diluent NS (ab193972)", "number":"AB213472-CMP13", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB Development Solution", "number":"AB213472-CMP06", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seals", "number":"AB213472-CMP11", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human IRAK4 Lyophilized Recombinant Protein", "number":"AB213472-CMP10", "productcode":"" }, { "size":"1 x 6 mL", "name":"Antibody Diluent 4BC", "number":"AB213472-CMP09", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human IRAK4 Capture Antibody", "number":"AB213472-CMP02", "productcode":"" }, { "size":"1 x 10 mL", "name":"5X Cell Extraction Buffer PTR (ab193970)", "number":"AB213472-CMP08", "productcode":"" }, { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB213472-CMP04", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human IRAK4 Detector Antibody", "number":"AB213472-CMP03", "productcode":"" } ] } } }

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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.

IRAK4 also known as Interleukin-1 receptor-associated kinase 4 is an important protein with an approximate molecular mass of 51 kDa. This protein is a significant part of the signaling cascade initiated by the Interleukin-1 receptor (IL-1R) and Toll-like receptors (TLRs). Expressed in a variety of tissues including the spleen thymus and lung IRAK4 is involved in innate immune responses. The presence of IRAK4 protein facilitates the downstream signaling pathways that activate the transcription factors responsible for inflammatory cytokine production.

Biological function summary

IRAK4 plays an important role in mediating signaling in the innate immune system by forming complexes with other proteins such as IRAK-1. These complexes propagate signals that result in the activation of transcription factors like NF-kB and AP-1 which regulate the expression of inflammatory genes. Through these mechanisms IRAK4 influences the body's ability to respond to infections and stresses. Its critical involvement in these signaling pathways highlights its importance in maintaining immune homeostasis.

Pathways

IRAK4 is a pivotal player in the Toll-like receptor and IL-1 receptor signaling pathways which are central to the host defense against pathogens. It functions upstream activating the MyD88-dependent pathway resulting in the recruitment and phosphorylation of other kinases such as IRAK-1 and TRAF6. These interactions trigger the NF-kB signaling cascade thereby enhancing pro-inflammatory cytokine production that is essential for initiating immune responses.

Mutations or dysregulation of IRAK4 are linked to increased susceptibility to recurrent bacterial infections and sepsis. Patients with IRAK4 deficiency have impaired NF-kB activation leading to a weakened inflammatory response. Additionally overexpression of IRAK4 has been observed in certain autoimmune diseases such as rheumatoid arthritis where the persistent activation of TLR signaling contributes to chronic inflammation. The role of IRAK4 in these disorders ties closely with its interaction with proteins like TRAF6 influencing disease development and progression.

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Product protocols

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Target data

Serine/threonine-protein kinase that plays a critical role in initiating innate immune response against foreign pathogens. Involved in Toll-like receptor (TLR) and IL-1R signaling pathways (PubMed : 17878374). Is rapidly recruited by MYD88 to the receptor-signaling complex upon TLR activation to form the Myddosome together with IRAK2. Phosphorylates initially IRAK1, thus stimulating the kinase activity and intensive autophosphorylation of IRAK1. Phosphorylates E3 ubiquitin ligases Pellino proteins (PELI1, PELI2 and PELI3) to promote pellino-mediated polyubiquitination of IRAK1. Then, the ubiquitin-binding domain of IKBKG/NEMO binds to polyubiquitinated IRAK1 bringing together the IRAK1-MAP3K7/TAK1-TRAF6 complex and the NEMO-IKKA-IKKB complex. In turn, MAP3K7/TAK1 activates IKKs (CHUK/IKKA and IKBKB/IKKB) leading to NF-kappa-B nuclear translocation and activation. Alternatively, phosphorylates TIRAP to promote its ubiquitination and subsequent degradation. Phosphorylates NCF1 and regulates NADPH oxidase activation after LPS stimulation suggesting a similar mechanism during microbial infections.

See full target information IRAK4

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websiteProtocolBooklet

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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Human IRAK4 ELISA Kit

ELISA - Human IRAK4 ELISA Kit (AB213472)

Functional Studies - Human IRAK4 ELISA Kit (AB213472)

Sandwich ELISA - Human IRAK4 ELISA Kit (AB213472)

Sandwich ELISA - Human IRAK4 ELISA Kit (AB213472)

Sandwich ELISA - Human IRAK4 ELISA Kit (AB213472)

Sandwich ELISA - Human IRAK4 ELISA Kit (AB213472)

Key facts

Detection method

Sample types

Reacts with

Assay type

Sensitivity

Range

Assay time

Assay Platform

Reactivity data

Product details

Precision

Recovery

What's included?

Properties and storage information

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Complementary Products

Assay Kits

Primary Antibodies

Secondary Antibodies

Assay Kits

Product promise