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IKK alpha Recombinant Rabbit Monoclonal Antibody [SN06-99]

Usd: 385

Specification

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Catalog# ET1611-15

IKK alpha Recombinant Rabbit Monoclonal Antibody [SN06-99]

Application

  • WB

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

IKK alpha Recombinant Rabbit Monoclonal Antibody [SN06-99]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within N-terminal human IKK alpha.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP, FC

Molecular Weight

Predicted band size: 85 kDa

Positive Control

HeLa cell lysate, HCT 116 cell lysate, SK-OV-3 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, rat kidney tissue, HeLa.

Conjugation

unconjugated

Clone Number

SN06-99

RRID

AB_3069992

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

  • 1:50-1:200

  • IP

  • Use at an assay dependent concentration.

  • FC

  • 1:1,000

Target

Function

Inhibitor of nuclear factor kappa-B kinase subunit alpha (IKK-α) also known as IKK1 or conserved helix-loop-helix ubiquitous kinase (CHUK) is a protein kinase that in humans is encoded by the CHUK gene. IKK-α is part of the IκB kinase complex that plays an important role in regulating the NF-κB transcription factor. However, IKK-α has many additional cellular targets, and is thought to function independently of the NF-κB pathway to regulate epidermal differentiation. IKK-α is a member of the serine/threonine protein kinase family and forms a complex in the cell with IKK-β and NEMO. IKK-α has been shown to function in epidermal differentiation independently of the NF-κB pathway. The zebrafish homolog of IKK-α has also been shown to play a role in the differentiation of the embryonic epithelium. IκB kinase α (IKKα) is a regulator of keratinocyte terminal differentiation and proliferation and plays a role in skin cancer.

Background References

1. Nielsen C et al. Immunodeficiency associated with a nonsense mutation of IKBKB. J Clin Immunol 34:916-21 (2014).

2. Thorne JL et al. Heterochromatin protein 1 gamma and IkB kinase alpha interdependence during tumour necrosis factor gene transcription elongation in activated macrophages. Nucleic Acids Res : (2012).

Sequence Similarity

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.

Tissue Specificity

Widely expressed.

Post-translational Modification

Phosphorylated by MAP3K14/NIK, AKT and to a lesser extent by MEKK1, and dephosphorylated by PP2A. Autophosphorylated.; (Microbial infection) Acetylation of Thr-179 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the I-kappa-B signaling pathway.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: O15111 Human

SwissProt: Q60680 Mouse

Entrez Gene:309361 Rat

Synonyms

chuk antibody

CHUK1 antibody

Conserved Helix Loop Helix Ubiquitous Kinase antibody

Conserved helix loop ubiquitous kinase antibody

Conserved helix-loop-helix ubiquitous kinase antibody

I Kappa B Kinase 1 antibody

I Kappa B Kinase Alpha antibody

I-kappa-B kinase 1 antibody

I-kappa-B kinase alpha antibody

IkappaB kinase antibody

Expand

Images

  • Western blot analysis of IKK alpha on different lysates with Rabbit anti-IKK alpha antibody (<a href="/products/ET1611-15" style="font-weight: bold;text-decoration: underline;">ET1611-15</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HCT 116 cell lysate (20 µg/Lane)<br />Lane 3: SK-OV-3 cell lysate (20 µg/Lane)<br />Lane 4: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 5: PC-12 cell lysate (20 µg/Lane)<br />Lane 6: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 7: Rat kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 85 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 17 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/ET1611-15" style="font-weight: bold;text-decoration: underline;">ET1611-15</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 IKK alpha on different lysates with Rabbit anti-IKK alpha antibody (ET1611-15) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HCT 116 cell lysate (20 µg/Lane)
    Lane 3: SK-OV-3 cell lysate (20 µg/Lane)
    Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 5: PC-12 cell lysate (20 µg/Lane)
    Lane 6: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 7: Rat kidney tissue lysate (40 µg/Lane)

    Predicted band size: 85 kDa
    Observed band size: 75 kDa

    Exposure time: 17 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 (ET1611-15) 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 rat kidney tissue with Rabbit anti-IKK alpha antibody (<a href="/products/ET1611-15" style="font-weight: bold;text-decoration: underline;">ET1611-15</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1611-15" style="font-weight: bold;text-decoration: underline;">ET1611-15</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 rat kidney tissue with Rabbit anti-IKK alpha antibody (ET1611-15) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1611-15) 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.

  • Flow cytometric analysis of HeLa cells labeling IKK alpha.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1611-15" style="font-weight: bold;text-decoration: underline;">ET1611-15</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling IKK alpha.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1611-15, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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

Citation

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IKK alpha Recombinant Rabbit Monoclonal Antibody [SN06-99] - BSA and Azide free

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