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Applications Table
| Western Blot | Chromatin IP | Immunoprecipitation |
|---|---|---|
| 1 - 2 µg/ml | — | 1 µg/ml |
for IkBa antibody (mAb)Background
IκBα – NFκB (NFκB p50 and NFκB p65) signaling is controlled to a large extent by the sequestration of the NFκB complex in the cytoplasm by its association with one of the IκB family of proteins. IκBα is phosphorylated at Ser32 and Ser36 (IκBα phospho Ser32,36) by the IκB Kinase (IKK) complex, resulting in the degradation of IκB and the nuclear translocation of NFκB.
for IkBa antibody (mAb)Immunogen
This antibody was raised against a recombinant protein corresponding to amino acid residues 32-291 of human IκBα.
for IkBa antibody (mAb)Application Notes
This IκBα antibody has been validated for use in Western blotting (1 - 2 µg/ml dilution) and immunoprecipitation (1 µg/ml dilution). For optimal results, primary antibody incubations should be performed at room temperature. The addition of 0.1% Tween 20 to all Blotto solutions may also reduce background. Individual optimization may be required.
for IkBa antibody (mAb)Buffer
PBS containing 0.02% sodium azide. Sodium azide is highly toxic.
for IkBa antibody (mAb)Positive Control
Active Motif's Jurkat nuclear extract (TPA + CI stimulated) (Catalog No. 36013) can be used as a positive control for IκBα antibody.
Bridging Antibody to Improve ChIP, MeDIP and IP
Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use IκBα antibody (mAb).
IκBα mAb tested by Western blot.
The anti-IκBα detects both phosphorylated and non-phosphorylated forms of IκBα by Western blot. Jurkat cells (~1 x 107) were treated for indicated time periods with 10 nm/ml PMA, 1 µM ionomycin and anti-CD28 (1:10,000 dilution). 10 µl of cytoplasmic (C) and nuclear extract (N) were resolved on a 8.75% SDS-PAGE and transferred to Immobilon membrane. Figure is courtesy of Dr. Shao-Cong Sun at Penn State Univ. College of Medicine.
for IkBa antibody (mAb)Storage
Antibodies in solution can be stored at -80°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for IkBa antibody (mAb)Guarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
Application Key
- ChIP = Chromatin Immunoprecipitation;
- DB = Dot Blot;
- ELISA = Enzyme-linked Immunosorbent Assay;
- EMSA = Electrophoretic Mobility Shift Assay
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
