GITR/TNFRSF18 Overexpression Lysate

Western Blot: GITR Overexpression Lysate (Adult Normal) [NBL1-17152] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GITR.

• Reactivity: Hu
• Applications: WB

GITR/TNFRSF18 Overexpression Lysate Summary

• Description:

  GITR/TNFRSF18 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC219997
  
  Accession#: NM_004195
  
  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: TNFRSF18

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: 23.5 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 GITR/TNFRSF18 Overexpression Lysate

• Activation-inducible TNFR family receptor
• AITR
• AITRTNF receptor superfamily activation-inducible protein
• CD357 antigen
• CD357
• GITR
• GITR-D
• GITRtumor necrosis factor receptor superfamily member 18
• Glucocorticoid-induced TNFR-related protein
• TNFRSF18
• tumor necrosis factor receptor superfamily, member 18

Background

The protein encoded by this gene is a member of the TNF-receptor superfamily. This receptor has been shown to have increased expression upon T-cell activation, and it is thought to play a key role in dominant immunological self-tolerance maintained by CD25(+)CD4(+) regulatory T cells. Knockout studies in mice also suggest the role of this receptor is in the regulation of CD3-driven T-cell activation and programmed cell death. Three alternatively spliced transcript variants of this gene encoding distinct isoforms have been reported. [provided by RefSeq]. Transcript Variant: This variant (1) represents the longest transcript. It contains an extra coding segment, which leads to a translation frame shift, compared to variant 2. The resulting isoform (1) contains a distinct and shorter C-terminus, as compared to isoform 2. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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: TNFRSF18