MSK1/RPS6KA5 Overexpression Lysate

Western Blot: RPS6KA5 Overexpression Lysate (Adult Normal) [NBL1-15572] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for RPS6KA5.

• Reactivity: Hu
• Applications: WB

MSK1/RPS6KA5 Overexpression Lysate Summary

• Description:

  MSK1/RPS6KA5 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200549
  
  Accession#: NM_182398
  
  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: RPS6KA5

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: 61.6 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 MSK1/RPS6KA5 Overexpression Lysate

• EC 2.7.11
• MGC1911,90 kDa ribosomal protein S6 kinase 5
• MSK1
• MSK1EC 2.7.11.1
• MSPK1
• Nuclear mitogen- and stress-activated protein kinase 1
• ribosomal protein S6 kinase alpha-5
• ribosomal protein S6 kinase, 90kD, polypeptide 5
• ribosomal protein S6 kinase, 90kDa, polypeptide 5
• RLPK
• RPS6KA5
• RSKL
• RSK-like protein kinase
• S6K-alpha-5

Background

MSK-1 is a mitogen and stress activated protein kinase-1 which belongs to the AGC family of kinases and is related in structure to the ribosomal p70 S6 kinase subfamily. MSK-1can be activated by ERK1/2 and SAPK2 /p38 MAP kinase. It is also known to be required for the phosphorylation of CREB, ATF1 H3 and HMG-14 in response to mitogen and stress (1-2). Similar to RSK, MSK-1 contains two kinase domains (N-term and a C-term) (3). Once phosphorylated on Thr581 and Ser360 by ERK1/2 and SAPK2/p38, MSK-1 autophosphorylate on at least 5 sites. Of these autophosphorylation sites Ser212 and Ser376 get phosphorylated by the C-terminal kinase domain of MSK-1 which is essential for the catalytic activity of the N-terminal kinase domain (4).

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: RPS6KA5