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IRAK4 Recombinant Rabbit Monoclonal Antibody [JE41-77]

Usd: 385

Specification

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Catalog# HA721347

IRAK4 Recombinant Rabbit Monoclonal Antibody [JE41-77]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Green monkey

Conjugation

  • unconjugated

Overview

Product Name

IRAK4 Recombinant Rabbit Monoclonal Antibody [JE41-77]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human IRAK4 aa 1-100 (N terminal).

Species Reactivity

Human, Mouse, Green monkey

Validated Applications

WB, IHC-P

Target Molecular Weight

Predicted band size: 52 kDa

Positive Control

Jurkat cell lysate, K-562 cell lysate, THP-1 cell lysate, RAW264.7 cell lysate, COS-1 cell lysate, HL-60 cell lysate, human skeletal muscle tissue.

Conjugation

unconjugated

Clone Number

JE41-77

RRID

AB_3072464

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:1,000

Target

Function

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. 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.

Background References

1. Somani VK et al. IRAK4 Signaling Drives Resistance to Checkpoint Immunotherapy in Pancreatic Ductal Adenocarcinoma. Gastroenterology. 2022 Jun

2. Bennett J et al. IRAK1 and IRAK4 as emerging therapeutic targets in hematologic malignancies. Curr Opin Hematol. 2022 Jan

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: Q9NWZ3 Human

SwissProt: Q8R4K2 Mouse

Synonyms

IL-1 receptor-associated kinase 4 antibody

Interleukin 1 receptor associated kinase 4 mutant form 1 antibody

Interleukin-1 receptor-associated kinase 4 antibody

Interleukin1 receptor associated kinase 4 antibody

IPD1 antibody

IRAK 4 antibody

IRAK-4 antibody

IRAK4 antibody

IRAK4 mutated form 1 antibody

IRAK4_HUMAN antibody

Expand

Images

  • Western blot analysis of IRAK4 on different lysates with Rabbit anti-IRAK4 antibody (<a href="/products/HA721347" style="font-weight: bold;text-decoration: underline;">HA721347</a>) at 1/1,000 dilution.<br /><br />Lane 1: Jurkat cell lysate<br />Lane 2: K-562 cell lysate<br />Lane 3: THP-1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 52 kDa<br />Observed band size: 52 kDa<br /><br />Exposure time: 22 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721347" style="font-weight: bold;text-decoration: underline;">HA721347</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of IRAK4 on different lysates with Rabbit anti-IRAK4 antibody (HA721347) at 1/1,000 dilution.

    Lane 1: Jurkat cell lysate
    Lane 2: K-562 cell lysate
    Lane 3: THP-1 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

    Exposure time: 22 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721347) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of IRAK4 on different lysates with Rabbit anti-IRAK4 antibody (<a href="/products/HA721347" style="font-weight: bold;text-decoration: underline;">HA721347</a>) at 1/1,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: COS-1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 52 kDa<br />Observed band size: 52 kDa<br /><br />Exposure time: 5 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721347" style="font-weight: bold;text-decoration: underline;">HA721347</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of IRAK4 on different lysates with Rabbit anti-IRAK4 antibody (HA721347) at 1/1,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: COS-1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

    Exposure time: 5 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721347) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of IRAK4 on different lysates with Rabbit anti-IRAK4 antibody (<a href="/products/HA721347" style="font-weight: bold;text-decoration: underline;">HA721347</a>) at 1/2,000 dilution.<br /><br />Lane 1: Jurkat cell lysate<br />Lane 2: THP-1 cell lysate<br />Lane 3: HL-60 cell lysate (low expression)<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 52 kDa<br />Observed band size: 52 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721347" style="font-weight: bold;text-decoration: underline;">HA721347</a>) at 1/2,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of IRAK4 on different lysates with Rabbit anti-IRAK4 antibody (HA721347) at 1/2,000 dilution.

    Lane 1: Jurkat cell lysate
    Lane 2: THP-1 cell lysate
    Lane 3: HL-60 cell lysate (low expression)

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

    Exposure time: 25 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721347) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-IRAK4 antibody (<a href="/products/HA721347" style="font-weight: bold;text-decoration: underline;">HA721347</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721347" style="font-weight: bold;text-decoration: underline;">HA721347</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-IRAK4 antibody (HA721347) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721347) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"