Myosin Phosphatase 2 Overexpression Lysate

Western Blot: Myosin Phosphatase 2 Overexpression Lysate (Adult Normal) [NBP2-06087] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Myosin Phosphatase 2.

• Reactivity: Hu
• Applications: WB

Myosin Phosphatase 2 Overexpression Lysate Summary

• Description:

  Myosin Phosphatase 2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC217905
  
  Accession#: NM_032104
  
  Protein Tag: C-MYC/DDK
  
  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: PPP1R12B

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: 26.1 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.

Alternate Names for Myosin Phosphatase 2 Overexpression Lysate

• MGC131980
• MGC87886
• Myosin phosphatase target subunit 2
• myosin phosphatase, target subunit 2
• Myosin phosphatase-targeting subunit 2
• MYPT 2
• MYPT2FLJ40942
• protein phosphatase 1 regulatory subunit 12B
• protein phosphatase 1, regulatory (inhibitor) subunit 12B

Background

PPP1R12B, also known as Protein phosphatase 1 regulatory subunit 12B, has 5 isoforms, a 982 amino acid isoform that is 110 kDa, a 386 amino acid isoform that is 43 kDa, a 224 amino acid isoform that is 26 kDa, a 208 amino acid isoform that is 24 kDa, and a 515 amino acid isoform that is 58 kDa; detected in skeletal muscle, fetal and adult heart, brain, placenta, kidney, spleen, thymus, pancreas and lung; with cytoplasmatic subcellular location; regulates myosin phosphatase activity and augments Ca(2+) sensitivity of the contractile apparatus. This protein is being studied for its involvement in breast cancer. The PPP1R12B protein has been shown to involve IL16, PPP1CB, PRKG1, HSPA8, MYL6, and MYL6B in RhoA pathway, PKA signaling, actin-based motility by Rho family GTPases, eIF2 pathway, CDK5 pathway, insulin pathway, G2/M transition, cell cycle, mitotic G2-G2/M phases, regulation of PLK1 activity at G2/M transition, cell cycle, mitotic, and vascular smooth muscle contraction pathways.

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: PPP1R12B