TIRAP (TLR2 and TLR4) Overexpression Lysate

Western Blot: TIRAP (TLR2 and TLR4) Overexpression Lysate (Native) [NBP2-08583] - TIRAP (TLR2 and TLR4) Overexpression Lysate (Native) Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for TIRAP.

• Reactivity: Hu
• Applications: WB

TIRAP (TLR2 and TLR4) Overexpression Lysate Summary

• Description:

  TIRAP (TLR2 and TLR4) Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC207162
  
  Accession#: NM_001039661
  
  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: TIRAP

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: 23.7 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 TIRAP (TLR2 and TLR4) Overexpression Lysate

• adapter protein wyatt
• Adaptor protein Wyatt
• FLJ42305
• Mal
• MyD88 adapter-like protein
• TIR domain-containing adapter protein
• toll/interleukin-1 receptor domain-containing adapter protein
• toll-interleukin 1 receptor (TIR) domain containing adaptor protein
• Toll-interleukin 1 receptor (TIR) domain-containing adaptor protein
• Toll-like receptor adaptor protein
• wyatt

Background

Mammalian Toll-like receptors (TLRs) recognize conserved products of microbial metabolism and activate NF-kappa B and other signaling pathways through the adapter protein MyD88. All TLRs have a Toll/IL-1 receptor (TIR) domain, which is responsible for signal transduction. MyD88 is one such protein that contains a TIR domain. It acts as an adapter, being involved in TLR-2, TLR-4 and TLR-9 signaling. Toll-interleukin 1 receptor (TIR) domain-containing adapter protein (TIRAP) controls activation of MyD88-independent signaling pathways downstream of TLR4. Double-stranded RNA-binding protein kinase PKR is a component of both the TIRAP- and MyD88-dependent signaling pathways (1). It has been demonstrated that TRIF associates with TRAF6 and TBK1 independently, and activates two distinct transcription factors, NF- B and IFN regulatory factor-3, respectively (2). TIRAP is differentially involved in signaling by members of the TLR family and may account for specificity in the downstream signaling of individual TLRs (3).

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: TIRAP