MLX (Max-like protein X; also known as transcription factor-like protein 4) is a 30kD member of the Max-like bHLHZip family of proteins. It is widely expressed and is found in both cytoplasm and nucleus. MLX is a transcriptional repressor when dimerized with MAD family members and a transcriptional activator when dimerized with MONDO family members. Human MLX is 298 amino acids (aa) in length. It contains one DNA-binding region (aa 76-87), an HLH domain (aa 127-192) and a Leu-zipper domain (aa 140-160). There are three alternate splice short forms. One shows the deletion of aa 15-68, a second shows deletion of aa 15-68 and 81-110, and a third shows a 3 aa substitution for aa 111-298. Over aa 197-298, human MLX is 99% identical to mouse and canine MLX.
Applications
Suitable for use in Western blot, Immunocytochemistry and ELISA. Other applications have not been tested.
Recommended Dilutions
Western blot: 0.1-0.2ug/ml. The detetction limit is ~2ng/lane under non-reducing and reducing conditions. Immunocytochemistry: 10ug/ml will detect human MLX in serum stimulated W138 human fetal lung fibroblasts. Fix cells with PBS containing 4% paraformaldehyde for 20 minutes at RT and block with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at RT. After blocking, incubate cells with diluted primary antibody for 3 hours at RT followed by NorthernLightsTM 557 donkey anti-goat lgG atRT for 1 hour. Between each step, wash cells with PBS containing 0.1% BSA. Direct ELISA: 0.5-1.0ug/ml. The detection limit is ~0.3ng/well. Optimal dilutions to be determined by the researcher.
Storage and Stability
Lyophilized and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store 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.
Immunogen
E. coli-derived recombinant human rhMLX (aa 197-298).
Form
Supplied as a lyophilized powder in PBS, 5% trehalose. Reconstitute with 500ul sterile PBS.
Purity
Purified by immunoaffinity chromatography.
Specificity
Recognizes human MLX. Species sequence homology: 99% identical to mouse and canine MLX.
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