PP2C alpha/PPM1A Overexpression Lysate

Western Blot: PPM1A Overexpression Lysate (Adult Normal) [NBL1-14656] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PPM1A.

• Reactivity: Hu
• Applications: WB

PP2C alpha/PPM1A Overexpression Lysate Summary

• Description:

  PP2C alpha/PPM1A Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC202070
  
  Accession#: NM_021003
  
  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: PPM1A

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: 42.3 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 PP2C alpha/PPM1A Overexpression Lysate

• EC 3.1.3.16
• FLJ42306
• MGC9201
• MMPa-2
• MPPa-1
• phosphatase 2C alpha
• PP2C alpha
• Pp2c1
• PP2CA
• PP2Calpha
• PP2C-ALPHA
• PPM1A
• PPPM1A
• protein phosphatase 1A (formerly 2C), magnesium-dependent, alpha isoform
• protein phosphatase 1A
• protein phosphatase 2C alpha isoform
• Protein phosphatase 2C isoform alpha
• Protein phosphatase IA
• protein phosphatase, Mg2+/Mn2+ dependent, 1A

Background

The protein encoded by this gene is a member of the PP2C family of Ser/Thr protein phosphatases. PP2C family members are known to be negative regulators of cell stress response pathways. This phosphatase dephosphorylates, and negatively regulates the activities of, MAP kinases and MAP kinase kinases. It has been shown to inhibit the activation of p38 and JNK kinase cascades induced by environmental stresses. This phosphatase can also dephosphorylate cyclin-dependent kinases, and thus may be involved in cell cycle control. Overexpression of this phosphatase is reported to activate the expression of the tumor suppressor gene TP53/p53, which leads to G2/M cell cycle arrest and apoptosis. Three alternatively spliced transcript variants encoding distinct isoforms have been described. [provided by RefSeq]. Transcript Variant: This variant (1) is the longest transcript and encodes isoform 1. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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: PPM1A