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Myosin Phosphatase Overexpression Lysate

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Western Blot: Myosin Phosphatase Overexpression Lysate (Adult Normal) [NBP2-11066] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Myosin Phosphatase.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Myosin Phosphatase Overexpression Lysate Summary

Description

Myosin Phosphatase Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC227493

Accession#: NM_001143885

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
PPP1R12A

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot.

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
115.1 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 Overexpression Lysate

  • MBSmyosin phosphatase, target subunit 1
  • MGC133042
  • Myosin phosphatase target subunit 1
  • Myosin phosphatase-targeting subunit 1
  • MYPT1protein phosphatase 1 regulatory subunit 12A
  • protein phosphatase 1, regulatory (inhibitor) subunit 12A
  • Protein phosphatase myosin-binding subunit

Background

The major protein phosphatase-1 (PP1) is composed of 3 subunits with apparent molecular mass of 110-130, 37 & 20 kDa. The 110-130 kDa component act as a regulatory subunit known as myosine binding (MBS) or myosine phosphatae targeting subunit (MYPT). The N-terminal portion of MYPT binds to both PP1c-delta and phosphorylated myosin, thereby increasing myosin phosphatase activity. Two isoform of MYPT have been identified (MYPT1 & MYPT2) and share 61% sequence identity. While MYPT1 is widely distributed in human tissues, MYPT2 is mostly detected in brain and heart.

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.

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Bioinformatics

Gene Symbol PPP1R12A