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PRODUCT CODE: ET1611-15

Recombinant IKK alpha Monoclonal Antibody (ET1611-15)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of IKK alpha on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-15, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br /> Positive control: <br /> Lane 1: Jurkat cell lysate<br /> Lane 2: Hela cell lysate
  • Western blot analysis of IKK alpha on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-15, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br /> Positive control: <br /> Lane 1: Jurkat cell lysate<br /> Lane 2: Hela cell lysate
  • ICC staining of IKK alpha in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-15, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded mouse skin tissue using anti-IKK alpha antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-15, 1/50) for 30 minutes 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 IKK alpha was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1611-15, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of IKK alpha on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-15, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Jurkat cell lysate
Lane 2: Hela cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant IKK alpha Monoclonal Antibody (ET1611-15)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Jurkat cell lysate, Hela cell lysate, SW480, mouse skin tissue, Hela.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SN06-99

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

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

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

85 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

O15111

PROTEIN NAME

IKK alpha

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; IkB kinase alpha subunit antibody; IkBKA antibody; IKK 1 antibody; IKK A antibody; IKK a kinase antibody; IKK-A antibody; IKK-alpha antibody; IKK1 antibody; IKKA antibody; IKKA_HUMAN antibody; Inhibitor Of Kappa Light Polypeptide Gene Enhancer In B Cells antibody; Inhibitor Of Nuclear Factor Kappa B Kinase Alpha Subunit antibody; Inhibitor of nuclear factor kappa-B kinase subunit alpha antibody; NFKBIKA antibody; Nuclear Factor Kappa B Inhibitor Kinase Alpha antibody; Nuclear factor NF kappa B inhibitor kinase alpha antibody; Nuclear factor NF-kappa-B inhibitor kinase alpha antibody; Nuclear factor NFkappaB inhibitor kinase alpha antibody; Nuclear Factor Of Kappa Light Chain Gene Enhancer In B Cells Inhibitor antibody; TCF-16 antibody; TCF16 antibody; Transcription factor 16 antibody

SEQUENCE SIMILARITIES

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.

FUNCTION

The transcription factor NFkB is retained in the cytoplasm in an inactive form by the inhibitory protein IkB. Activation of NFkB requires that IkB be phosphorylated on specific serine residues, which results in targeted degradation of IkB. IkB kinase a (IKKa), previously designated CHUK, interacts with IkB-a and specifically phosphorylates IkB-a on Ser 32 and 36, the sites that trigger its degradation. IKKa appears to be critical for NFkB activation in response to proinflammatory cytokines. Phosphorylation of IkB by IKKa is stimulated by the NFkB inducing kinase (NIK), which itself is a central regulator for NFkB activation in response to TNF and IL-1. The functional IKK complex contains three subunits, IKKa, IKKb and IKKg (also designated NEMO), and each appear to make essential contributions to IkB phosphorylation.