IKK epsilon Antibody

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ProSci
Product Code: PSI-2329
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-2329-0.02mg0.02mg£150.00
Quantity:
PSI-2329-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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<strong>Figure 1 Induction Validation in Mouse Cell Line</strong><br> Loading: 15 μg of Raw264.7 cell lysates per lane. Antibodies: IKK epsilon 2329 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Cells were treated with LPS (0.3 μg/mL) for 3 hrs (lane 2) and 6 hrs (lane 3) or with control (lane 1).
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<strong>Figure 2 Western Blot Validation in Human and Mouse Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IKK epsilon 2329 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 3 Western Blot Validation in Mouse Tissue Lysates</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IKK epsilon 2329 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 4 Western Blot Validation in Jurkat Cell Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IKK epsilon 2329 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 5 Immunohistochemistry Validation of IKK epsilon in Human Pancreas Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-IKK epsilon antibody (2329) at 10 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 6 Immunofluorescence Validation of IKK epsilon in HeLa Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling IKK epsilon with 2329 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
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<strong>Figure 1 Induction Validation in Mouse Cell Line</strong><br> Loading: 15 μg of Raw264.7 cell lysates per lane. Antibodies: IKK epsilon 2329 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Cells were treated with LPS (0.3 μg/mL) for 3 hrs (lane 2) and 6 hrs (lane 3) or with control (lane 1).
<strong>Figure 2 Western Blot Validation in Human and Mouse Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IKK epsilon 2329 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Western Blot Validation in Mouse Tissue Lysates</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IKK epsilon 2329 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Western Blot Validation in Jurkat Cell Lysate</strong><br> Loading: 15 μg of lysates per lane. Antibodies: IKK epsilon 2329 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 5 Immunohistochemistry Validation of IKK epsilon in Human Pancreas Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-IKK epsilon antibody (2329) at 10 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 6 Immunofluorescence Validation of IKK epsilon in HeLa Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling IKK epsilon with 2329 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Documents

Further Information

Additional Names:
IKK epsilon Antibody: IKKE, IKKI, IKK-E, IKK-i, IKKE, KIAA0151, Inhibitor of nuclear factor kappa-B kinase subunit epsilon, Inducible I kappa-B kinase, I-kappa-B kinase epsilon
Application Note:
WB: 1-2 μg/mL; IHC: 10 μg/mL; IF: 20 μg/mL.

Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in human samples; Immunofluorescence in human samples. All other applications and species not yet tested.
Background:
IKK epsilon Antibody: Nuclear factor kappa B (NF-κB) is a ubiquitous transcription factor and an essential mediator of gene expression during activation of immune and inflammatory responses. NF-κB mediates the expression of a great variety of genes in response to extracellular stimuli. NF-κB is associated with IκB proteins in the cell cytoplasm, which inhibit NF-κB activity. IκB is phosphorylated by IκB kinase (IKK) complex that contains IKKα, IKKβ, and IKKγ. A novel molecule in the IKK complex was recently identified and designated IKKε/IKK-i. IKK epsilon is required for the activation of NF-κB by PMA and T cell receptors but not by TNFα and IL-1. IKKε/IKK-i message is expressed in a variety of tissues and is inducible by TNFα, IL-1, and LPS.
Background References:
  • Peters et al. Mol Cell 2000;5(3):513-22.
  • Shimada et al. Int Immunol 1999;11(8):1357-62 (WD0101).
Buffer:
IKK epsilon Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/ml
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Homology:
Predicted species reactivity based on immunogen sequence: Rat: (75%)
Immunogen:
Anti-IKK epsilon antibody (2329) was raised against a peptide corresponding to 16 amino acids near the carboxy terminus of human IKK epsilon.

The immunogen is located within the last 50 amino acids of IKK epsilon.
ISOFORMS:
Human IKK epsilon has 2 isoforms, including isoform 1 (716aa, 80kD) and isoform 2 (631aa, 70kD). Mouse IKK epsilon has one isoform (717aa, 81kD) and Rat IKK epsilon also has one isoform (717aa, 80kD). 2329 can detect human, mouse and possibly rat.
NCBI Gene ID #:
9641
NCBI Official Name:
inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase epsilon
NCBI Official Symbol:
IKBKE
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 80kD

Observed: 80 kD
Protein Accession #:
AF241789
Protein GI Number:
7288877
Purification:
IKK epsilon Antibody is affinity chromatography purified via peptide column.
Research Area:
Signal Transduction
SPECIFICITY:
It has no cross response to IKKa, IKKb, or IKKy.
Swissprot #:
Q14164
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

Induced Expression Validation (Figure 1): Raw 264.7 cells were treated with LPS (0.3 µg /mL) for different time period (3 hrs and 6 hrs). IKK epsilon expression detected by anit-IKK epsilon antibodies (2329) was induced by LPS treatment and the expression levels were increased gradually at 3 hrs and 6 hrs.

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