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IRAK-1 Recombinant Rabbit Monoclonal Antibody [JE60-03]

Usd: 385

Specification

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Catalog# HA720079

IRAK-1 Recombinant Rabbit Monoclonal Antibody [JE60-03]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

Overview

Product Name

IRAK-1 Recombinant Rabbit Monoclonal Antibody [JE60-03]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human IRAK-1 aa 201-500/712.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 77 kDa

Positive Control

A549 cell lysate, HeLa cell lysate, human colon cancer tissue.

Conjugation

unconjugated

Clone Number

JE60-03

RRID

AB_3072018

Product Features

Form

Liquid

Concentration

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

  • 1:2,000

  • IHC-P

  • 1:100-1:200

  • IP

  • 1-2μg/sample

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. Association with MYD88 leads to IRAK1 phosphorylation by IRAK4 and subsequent autophosphorylation and kinase activation. 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 the interferon regulatory factor 7 (IRF7) to induce its activation and translocation to the nucleus, resulting in transcriptional activation of type I IFN genes, which drive the cell in an antiviral state. When sumoylated, translocates to the nucleus and phosphorylates STAT3.

Background References

1. Liu X. et. al. BAP31 regulates IRAK1-dependent neuroinflammation in microglia. J Neuroinflammation. 2019 Dec

2. Liu PH. et. al. An IRAK1-PIN1 signalling axis drives intrinsic tumour resistance to radiation therapy. Nat Cell Biol. 2019 Feb

Subcellular Location

Cytoplasm, Lipid droplet, Nucleus.

UNIPROT

SwissProt: P51617 Human

SwissProt: Q62406 Mouse

Synonyms

AA48924 antibody

Il1rak antibody

Interleukin 1 receptor associated kinase 1 antibody

Interleukin-1 receptor-associated kinase 1 antibody

IRAK antibody

IRAK-1 antibody

Irak1 antibody

IRAK1-S antibody

IRAK1_HUMAN antibody

mPLK antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of IRAK-1 on different lysates with Rabbit anti-IRAK-1 antibody (<a href="/products/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si IRAK-1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 77 kDa<br />Observed band size: 75/77 kDa<br /><br />Exposure time: 3 minutes; 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/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a>) at 1/2,000 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.

    ☑ Knockdown (KD)

    Western blot analysis of IRAK-1 on different lysates with Rabbit anti-IRAK-1 antibody (HA720079) at 1/2,000 dilution.

    Lane 1: A549-si NT cell lysate
    Lane 2: A549-si IRAK-1 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 77 kDa
    Observed band size: 75/77 kDa

    Exposure time: 3 minutes; 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 (HA720079) at 1/2,000 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.

  • Western blot analysis of IRAK-1 on different lysates with Rabbit anti-IRAK-1 antibody (<a href="/products/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549 cell lysate<br />Lane 2: HeLa cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 77 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 30 seconds; ECL: K1802;<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/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a>) at 1/2,000 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 IRAK-1 on different lysates with Rabbit anti-IRAK-1 antibody (HA720079) at 1/2,000 dilution.

    Lane 1: A549 cell lysate
    Lane 2: HeLa cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 77 kDa
    Observed band size: 75 kDa

    Exposure time: 30 seconds; ECL: K1802;

    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 (HA720079) at 1/2,000 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.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-IRAK-1 antibody (<a href="/products/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a>) at 1/200 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/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a>) at 1/200 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 colon cancer tissue with Rabbit anti-IRAK-1 antibody (HA720079) at 1/200 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 (HA720079) at 1/200 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.

  • IRAK-1 was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a> at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA720079" style="font-weight: bold;text-decoration: underline;">HA720079</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    IRAK-1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA720079 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA720079 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA720079 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA720079 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute; ECL: K1802

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

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metafield image

IRAK-1 Rabbit Polyclonal Antibody

Application: WB,IHC-P,IF-Cell

Reactivity: Human,Mouse

Conjugate: unconjugated