		Technical Data
A0750-10	ACT1 (NF-kB Activator 1, Nuclear Factor kB Activator 1)

Description:
NF-kB (nuclear factor kB) is sequestered in the cytoplasm by IkB family of inhibitory proteins that mask the nuclear localization signal of NF-kB thereby preventing translocation of NF-kB to the nucleus (1). External stimuli such as tumor necrosis factor or other cytokines, such as IL-1 results in phosphorylation and degradation of IkB releasing NF-kB dimers. NF-kB dimer subsequently translocates to the nucleus and activates target genes (2). Recently, Li et al. (3) have isolated a cDNA coding for a protein named Act1 (NF-kB activator 1). Act1 cDNA codes for a protein containing 574 aa with a predicated molecular mass of 60kD. Act1 protein contains a helix-loop-helix domain in its N-terminal portion and a coiled-coil structure at the C terminus. Act1 lacks any catalytic domain, however, it activates IKK through the helix-loop-helix domain. The activation of NF-kB by Act1 appears to be through the NIK-IKK complex, because a NIK dominant-negative mutant protein inhibited Act1-induced NF-kB activation. IL-1 responsive pathway is also necessary for Act1 function. Act1 mRNA is expressed in a variety of tissues, with high level in thymus, kidney and placenta; moderate level in heart, skeletal muscle, colon, liver, lung and small intestine; and at a very low level in brain, spleen, and peripheral blood leukocytes.

Applications:
Suitable for use in Western Blot. Other applications not tested.

Recommended Dilution:
Western Blot: 1-2ug/ml. Whole cell lysate from a breast cancer cell line, MCF-7, can be used as positive control and a protein of approximate molecular weight of 60kD band is detected.
Optimal dilutions to be determined by the researcher.

Recommended Positive Control:
C2069-44H1: MCF-7 Cell Lysate

Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile glycerol (40-50%), aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Type	Isotype	Clone	Grade
Pab	IgG		Affinity Purified

Size		Storage	Shipping	Source	Host
100ug		4°C (-20°C Glycerol)	Blue Ice	Human	Rabbit

Concentration:

~0.5mg/ml

Immunogen:

A mixture of synthetic peptides corresponding to aa341-351, 91-104 and 562-574 of human ACT1.

Purity:

Purified by Protein G affinity chromatography.

Form

Supplied as a liquid in PBS, pH 7.2, 0.02% sodium azide.

Specificity:

Recognizes human ACT1.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

1. Verma, I.M., et al., Genes. Dev. 9: 2723-2735 (1995). 2. Verma, I. & Stevenson, J.K., Proc. Nat. Acad. Sci. USA 94: 11,758 (1997). 3. Li, X., et al., Proc. Natl. Acad. Sci. USA 97(19): 10,489-10,493 (2000).