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PIASy Overexpression Lysate

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Western Blot: PIASy Overexpression Lysate (Adult Normal) [NBL1-14388] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PIASy.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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PIASy Overexpression Lysate Summary

Description

PIASy Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC206748

Accession#: NM_015897

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
PIAS4

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
56.3 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.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

Alternate Names for PIASy Overexpression Lysate

  • E3 SUMO-protein ligase PIAS4
  • FLJ12419
  • MGC35296
  • Piasg
  • PIAS-gamma
  • PIASY
  • Protein inhibitor of activated STAT protein 4
  • Protein inhibitor of activated STAT protein gamma
  • protein inhibitor of activated STAT protein PIASy
  • protein inhibitor of activated STAT, 4
  • zinc finger, MIZ-type containing 6
  • ZMIZ6

Background

The IL-6-type family of cytokines, which includes IL-6 as well as a number of similar cytokines and growth factors, plays a significant role in regulating gene activation, proliferation and differentiation. Transcription factors of the Stat family are known to be involved in this signal transduction pathway, undergoing phosphorylation, dimerization and translocation to the nucleus upon activation. PIAS 1, for protein inhibitor of activated Stat1 (also designated Gu/RNA helicase II binding protein), binds specifically to Stat1, blocking Stat1 DNA-binding activity and inhibiting Stat1-mediated gene activation. PIAS 1 also binds to the Gu/RNA helicase II enzyme, leading to the proteolytic cleavage of Gu/RH-II. PIAS 3 similarly binds specifically to Stat3, blocking Stat3 DNA-binding activity and inhibiting Stat3-mediated gene activation.

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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Product General Protocols

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Video Protocols

WB Video Protocol

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Bioinformatics

Gene Symbol PIAS4