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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).
Figure 1 Induction Validation in Mouse Cell Line
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).
Cell Treatment
Trial Size

IKK epsilon Antibody

CATALOG NUMBER: 2329

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
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.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
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.
Specificity:
It has no cross response to IKKa, IKKb, or IKKy.
Positive Control 1:
Cat. No. 1205 - Jurkat Cell Lysate
Positive Control 2:
Cat. No. 1307 - Human Pancreas Tissue Lysate
Positive Control 3:
Cat. No. 11-101 - Human Pancreas Tissue Slide
Positive Control 4:
Cat. No. 1202 - A431 Cell Lysate
Positive Control 5:
Cat. No. 1224 - Daudi Cell Lysate
Predicted Molecular Weight:
Predicted: 80kD
Observed: 80 kD
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.

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.
Purification:
IKK epsilon Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
IKK epsilon Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/ml
Storage Conditions:
IKK epsilon antibody can be stored at 4°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
IKBKE
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
Protein Accession Number:
AF241789
Protein Gi Number:
7288877
Ncbi Gene Id Number:
9641
User Note:
Optimal dilutions for each application to be determined by the researcher.
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 Reference 1:
Peters et al. Mol Cell 2000;5(3):513-22.
Background Reference 2:
Shimada et al. Int Immunol 1999;11(8):1357-62 (WD0101).

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

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.

CATALOG NUMBER:

2329

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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