MLX Overexpression Lysate

Western Blot: MLX Overexpression Lysate (Adult Normal) [NBP2-05216] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for MLX.

Product Discontinued

MLX Overexpression Lysate Summary

• Description: Transient overexpression lysate of MAX-like protein X (MLX), transcript variant 3 The lysate was created in HEK293T cells, using Plasmid ID RC224283 and based on accession number NM_170607. The protein contains a C-MYC/DDK Tag. You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: MLX

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 33.1 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

Alternate Names for MLX Overexpression Lysate

• BHLHD13
• bHLHd13Max-like bHLHZip protein
• BigMax alpha
• BigMax protein
• Class D basic helix-loop-helix protein 13
• MAD7
• MAX-like protein X
• MLX
• MXD7
• TCFL4
• TCFL4Protein BigMax
• transcription factor-like 4
• Transcription factor-like protein 4

Background

Max is a nuclear localized bHLH-Zip protein that forms homodimers or heterodimers with Myc family members, including Myc, Mad 1, Mad 3, Mad 4, Mxi1 and Mnt (or Rox). These dimers bind to the E-box sequence CACGTG in order to regulate cell growth, proliferation and apoptosis. Mlx (Max-like protein X) is a bHLH-Zip protein that is structurally and functionally related to Max. Like Max, Mlx is broadly expressed in many tissues and has a long half-life. Mlx also forms homodimers or heterodimers with members of the Myc family, specifically Mad 1, Mad 4 and Rox, and members of the Mondo family, to repress or activate transcription from CACGTG E-boxes. MondoA forms weak homodimers and preferentially forms heterodimers with Mlx. The MondoA/Mlx complex is primarily localized to the cytoplasm, but will translocate to the nucleus in response to leptomycin B. Mlx can also dimerize with WBSCR14, a protein involved in Williams-Beuren syndrome (WBS), to repress E-box transcription, which provides further evidence that Mlx is a critical element in a transcription factor network.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: MLX