CD74 Overexpression Lysate

Western Blot: CD74 Overexpression Lysate (Adult Normal) [NBL1-08968] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD74.

• Reactivity: Hu
• Applications: WB

CD74 Overexpression Lysate Summary

• Description:

  CD74 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC212964
  
  Accession#: NM_001025159
  
  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: CD74

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: 33.3 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 CD74 Overexpression Lysate

• CD74 antigen (invariant polypeptide of major histocompatibility complex, classII antigen-associated)
• CD74 antigen
• CD74 molecule, major histocompatibility complex, class II invariant chain
• CD74
• CLIP
• DHLAG
• DHLAGgamma chain of class II antigens
• HLA class II histocompatibility antigen gamma chain
• HLADG
• HLA-DR antigens-associated invariant chain
• HLA-DR-gamma
• Ia antigen-associated invariant chain
• Ia-associated invariant chain
• Ia-gamma
• Ii
• INVG34
• MHC HLA-DR gamma chain
• p33

Background

CD74, also known as the MHC class II associated invariant chain (Ii), is a type II transmembrane protein which binds to the peptide binding groove of newly synthesized MHC class II alpha/beta heterodimers and prevents their premature association with endogenous polypeptides. CD74 is produced in molar excess over MHC class II and some of this is expressed by an unknown pathway on the cell surface independent of, or in association with, MHC class II molecules. The half life of CD74 on the cell surface is only 3 to 4 min after which it is internalized. It has been proposed that cell surface CD74 binds MHC class II molecules which have lost their antigenic peptide leading to internalization and reloading with antigenic peptide. The transport route of class II alpha/beta/Ii complexes is regulated selectively by two forms of CD74 (p33 and p35) which are generated by the use of alternative translation initiation sites. CD74 is expressed primarily by antigen presenting cells such as B lymphocytes (from before the pre-B cell stage to before the plasma cell stage), macrophages and monocytes together, with many epithelial cells. CD74 may exist in different isoforms ranging in size from 33 to 41 kDa, depending on genetic splicing.

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: CD74