IKBKE Recombinant Rabbit Monoclonal Antibody [JU06-72]
Catalog# ET1706-20
IKBKE Recombinant Rabbit Monoclonal Antibody [JU06-72]
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WB
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IF-Cell
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FC
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IHC-P
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Human
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unconjugated
Overview
Product Name
IKBKE Recombinant Rabbit Monoclonal Antibody [JU06-72]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human IKBKE aa 26-75 / 716.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, FC, IHC-P
Molecular Weight
Predicted band size: 80 kDa
Positive Control
Human spleen tissue, human thymus tissue, HeLa cell lysate, Raji cell lysate, Daudi cell lysate, Hela, JAR, SK-Br-3.
Conjugation
unconjugated
Clone Number
JU06-72
RRID
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
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WB
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1:500-1:2,000
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IF-Cell
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1:50-1:200
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FC
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1:50-1:100
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IHC-P
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1:50-1:200
Target
Function
The transcription factor NFκB is retained in the cytoplasm in an inactive form by the inhibitory protein IκB. Activation of NFκB requires that IκB be phosphorylated on specific serine residues, which results in targeted degradation of IκB. IκB kinase α (IKKα), previously designated CHUK, interacts with IκB-α and specifically phosphorylates IκB-α on the sites that trigger its degradation, Serines 32 and 36. The functional IKK complex contains three subunits, IKKα, IKKβ and IKKγ (also designated NEMO), and each appear to make essential contributions to IκB phosphorylation. IKK-i is a serine/threonine kinase that shares homology with IKKα and IKKβ. IKK-i is primarily expressed in immune cells and is induced by lipopolysaccharide and by proinflammatory cytokines including TNFα, IL-1 and IL-6. Overexpression of IKK-i has been shown to result in phosphorylation of IκBα on Ser 32 and Ser 36, and in NFκB activation, suggesting that IKK-i may act as an IκB kinase in the immune system.
Background References
1. Gu L et al. Human DEAD box helicase 3 couples I B kinase e to interferon regulatory factor 3 activation. Mol Cell Biol 33:2004-15 (2013).
2. Paladino P et al. Cellular localization of the herpes simplex virus ICP0 protein dictates its ability to block IRF3-mediated innate immune responses. PLoS One 5:e10428 (2010).
Sequence Similarity
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.
Tissue Specificity
Highly expressed in spleen followed by thymus, peripheral blood leukocytes, pancreas, placenta. Weakly expressed in lung, kidney, prostate, ovary and colon.
Post-translational Modification
Autophosphorylated and phosphorylated by IKBKB/IKKB. Phosphorylation at Ser-172 is enhanced by the interaction with DDX3X. Phosphorylated at Thr-501 upon IFN activation.; Sumoylation by TOPORS upon DNA damage is required for protection of cells against DNA damage-induced cell death. Desumoylated by SENP1.; 'Lys-63'-linked polyubiquitinated at Lys-30 and Lys-401 by TRAF2:BIRC2 and TRAF2:BIRC3 complexes. Ubiquitination is induced by LPS, TNFA and interleukin-1 and required for full kinase activity and KF-kappa-B pathway activation.
Subcellular Location
Cytoplasm, Nucleus.
UNIPROT
Synonyms
I kappa B kinase epsilon antibody
I-kappa-B kinase epsilon antibody
IkBKE antibody
IKK related kinase epsilon antibody
IKK-E antibody
IKK-epsilon antibody
IKK-i antibody
IKKE antibody
IKKE_HUMAN antibody
IKKepsilon antibody
ExpandImages
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Western blot analysis of IKBKE on different lysates with Rabbit anti-IKBKE antibody (ET1706-20) at 1/2,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: Raji cell lysate (20 µg/Lane)
Lane 3: Daudi cell lysate (20 µg/Lane)
Predicted band size: 80 kDa
Observed band size: 75 kDa
Exposure time: Lane 1: 3 minutes; Lane 2-3: 30 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 (ET1706-20) 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 spleen tissue with Rabbit anti-IKBKE antibody (ET1706-20) 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 (ET1706-20) 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 thymus tissue with Rabbit anti-IKBKE antibody (ET1706-20) 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 (ET1706-20) 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. -
Immunocytochemistry analysis of Hela cells labeling IKBKE with Rabbit anti-IKBKE antibody (ET1706-20) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-IKBKE antibody (ET1706-20) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilutionNuclear DNA was labelled in blue with DAPI. -
Immunocytochemistry analysis of JAR cells labeling IKBKE with Rabbit anti-IKBKE antibody (ET1706-20) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-IKBKE antibody (ET1706-20) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. -
Immunocytochemistry analysis of SK-Br-3 cells labeling IKBKE with Rabbit anti-IKBKE antibody (ET1706-20) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-IKBKE antibody (ET1706-20) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"