抗IkB-alpha (6A920)抗体 | Anti-IkB-alpha (6A920) Antibody

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	Western Blot: IkB-alpha Antibody (6A920) [NB100-56507] - Lysates of Jurkat human acute T cell leukemia cell line, LNCaP human prostate cancer cell line, PCx2011;3 human prostate cancer cell line, HeLa human cervical epithelial carcinoma cell line, and NIHx2011;3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 0.5 ug/mL mouse anti-IkB-a monoclonal (NB100-56507, Novus Biologicals), followed by 1:2000 dilution of the appropriate HRP-conjugated secondary antibody, donkey anti-mouse IgG (HAF018).
	Immunohistochemistry-Paraffin: IkB-alpha Antibody (6A920) [NB100-56507] - Formalin-fixed, paraffin-embedded human breast probed with IkBa antibody at 5 ug/ml. Human tissue TMA was used for this test. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min..
	Flow Cytometry: IkB-alpha Antibody (6A920) [NB100-56507] - Intracellular staining of 10^6 ThP-1 cells using 0.25 ug of NB100-56507. Shaded histogram represents cells alone, green represents the isotype control, and red represents the IkBa antibody. Novus's intracellular flow kit was used for this test, and an anti-mouse IgG FITC conjugated secondary.
	Western Blot: IkB-alpha Antibody (6A920) [NB100-56507] - IkBa using NB100-56507 at 2 ug/ml in (A) Daudi and (B) NIH 3T3 whole cell lysate.
	Biological Strategies Validation. Western Blot: IkB-alpha Antibody (6A920) [NB100-56507] - Jurkat cells were treated for 30 min with 100 ug/ml ALLN (N-Acetyl-Leu-Leu-Norleucinal; a Calpain inhibitor and also proteasome inhibitor that prevents IkBa dephosphorylation) followed by incubation with (lanes 2 & 4) or without 1 nM TNF-a (1 & 3). The membranes were blotted with NB100-56724 (lanes 1 & 2) or NB100-56507 (that recognizes both non-phospho and phosphorylated forms of IkBa) and immunoreactivity was detected by ECL. The data shows that NB100-56724 detects specifically the phosphorylated form of IkBa.
	Simple Western: IkB-alpha Antibody (6A920) [NB100-56507] - Simple Western lane view shows a specific band for IKB alpha in 0.5 mg/ml of NIH-3T3 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

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Background

NF-kB is silenced in the cytoplasm by an inhibitory protein, IkB (1). Synthesis of IkBa is autoregulated (2). IkB proteins are phosphorylated by IkB kinase complex consisting of at least three proteins, IKK1/a, IKK2/b, and IKK3/g (3-6). External stimuli such as tumor necrosis factor or other cytokines results in phosphorylation and degradation of IkB releasing NF-kB dimers. NF-kB dimer subsequently translocates to the nucleus and activates target genes. Six members of IkB family members have been identified (1). One of the first gene induced following NF-kB activation is IkBa.

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