TPT1/TCTP Overexpression Lysate

Western Blot: TCTP Overexpression Lysate (Adult Normal) [NBL1-17234] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for TCTP.

• Reactivity: Hu
• Applications: WB

TPT1/TCTP Overexpression Lysate Summary

• Description:

  TPT1/TCTP Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC201664
  
  Accession#: NM_003295
  
  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: TPT1

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: 19.4 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 TPT1/TCTP Overexpression Lysate

• FLJ27337
• Fortilin
• Histamine-releasing factor
• HRF
• p02
• p23
• TCTP
• TCTPHRFfortilin
• TPT1
• translationally-controlled tumor protein
• tumor protein, translationally-controlled 1

Background

Histamine is an inflammatory mediator that is ubiquitously expressed and has a broad range of pharmacologic effects. Specifically, it plays a role in the central nervous, gastrointestinal, respiratory and immune systems. Histamine release is mediated by the stimulation of mast cells and basophils. Histamine-releasing factor (HRF) is a cytokine-like molecule that causes the release of histamine, IL-4 and IL-13 from basophils as well as the secretion of IL-8 and a calcium response in eosinophils. HRF belongs to the translationally controlled tumor protein (TCTP) family and is detected as a protein of 21 kDa in mice and 23 kDa in human. It is expressed in several healthy and tumoral cells, including erythrocytes, hepatocytes, macrophages, platelets, keratinocytes, erythroleukemia cells, gliomas, melanomas, hepatoblastomas and lymphomas, and it is localized in the cytoplasm. HRF plays a pivotal role in allergic diseases, and due to its wide distribution in brain, is thought to be involved in neurodegenerative disorders, such as Alzheimer's disease and Down Syndrome.

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: TPT1