BTK Overexpression Lysate

Western Blot: BTK Overexpression Lysate (Adult Normal) [NBP2-09059] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for BTK.

• Reactivity: Hu
• Applications: WB

BTK Overexpression Lysate Summary

• Description:

  BTK Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC211582
  
  Accession#: NM_000061
  
  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: BTK

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: 76.1 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 BTK Overexpression Lysate

• Agammaglobulinaemia tyrosine kinase
• AGMX1
• AGMX1MGC126262
• AT
• ATKIMD1
• B-cell progenitor kinase
• BPK
• Bruton agammaglobulinemia tyrosine kinase
• Bruton tyrosine kinase
• BTK
• dominant-negative kinase-deficient Brutons tyrosine kinase
• EC 2.7.10
• EC 2.7.10.2
• IMD1
• PSCTK1
• tyrosine-protein kinase BTK
• XLA
• XLAMGC126261

Background

Brutons tyrosine kinase (BTK) is a member of the BTK/Tec family of cytoplasmic tyrosine kinases. All members of the BTK/Tec family contain SH3 and SH2 domains and, with the exception of Txk and Dsrc28C, also contain a pleckstrin homology (PH) and a Tec homology (TH) domain in their amino termini. BTK plays an important role in B cell development. Activation of B cells by various ligands is accompanied by BTK membrane translocation mediated by its PH domain binding to phosphatidylinositol-3,4,5-trisphosphate. The membrane located BTK is active and associated with transient phosphorylation of two tyrosine residues, Tyr551 and Tyr223. Tyr551 in the activation loop is transphosphorylated by the Src family tyrosine kinase, leading to autophosphorylation at Tyr223 within the SH3 domain, which is necessary for full 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.

Bioinformatics

• Gene Symbol: BTK