CD5 Overexpression Lysate

Western Blot: CD5 Overexpression Lysate (Adult Normal) [NBL1-08956] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD5.

• Reactivity: Hu
• Applications: WB

CD5 Overexpression Lysate Summary

• Description:

  CD5 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC206494
  
  Accession#: NM_014207
  
  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: CD5

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: 54.4 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 CD5 Overexpression Lysate

• CD5 antigen (p56-62)
• CD5 antigen
• CD5 molecule
• CD5
• LEU1T-cell surface glycoprotein CD5
• Lymphocyte antigen T1/Leu-1
• T1

Background

In humans, CD5 is a 55kDa T lymphocyte single chain transmembrane glycoprotein. It is present on all mature T lymphocytes, on most thymocytes and on many T cell leukemias and lymphomas. It reacts with a subpopulation of activated B cells. CD5/Lyt1 antigen is a monomeric type I transmembrane glycoprotein expressed on thymocytes, T lymphocytes, and a subset of B lymphocytes, but not on natural killer (NK) cells. It has been identified as the major ligand of the B cell antigen CD72. The frequency of CD5+ B cells exhibits strain dependent variation, and the phenotypic, anatomical, functional, developmental, and pathological characteristics of the CD5+ B cells suggest that they may represent a distinct lineage, known as B1 cells. Binding of CD5 on the T cell surface can augment alloantigen or mitogen induced lymphocyte proliferation and induces increased cytosolic free calcium, IL2 secretion, and IL2R expression. It has been proposed that CD5 negatively regulates signal transduction mediated by the T cell and B cell receptors.

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: CD5