BAG5 Overexpression Lysate

Western Blot: BAG5 Overexpression Lysate (Adult Normal) [NBP2-09157] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for BAG5.

• Reactivity: Hu
• Applications: WB

BAG5 Overexpression Lysate Summary

• Description:

  BAG5 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC212765
  
  Accession#: NM_001015048
  
  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: BAG5

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: 51 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 BAG5 Overexpression Lysate

• BAG family molecular chaperone regulator 5
• BAG-5
• BAG-family molecular chaperone regulator-5
• Bcl-2-associated athanogene 5
• BCL2-associated athanogene 5
• KIAA0873

Background

The BAG (Bcl-2-associated anthanogene) proteins are a family of chaperone regulators that modulate a number of diverse processes including proliferation, survival, stress responses, tumorigenesis, neuronal differentiation, growth arrest and apoptosis (reviewed Takayama and Reed, 2001; Doong et al, 2002, and Doukhanina et al. 2006). BAG proteins have been characterized as co-chaperones and interact with the chaperone heat shock proteins 70, both constitutive Hsc70 and inducible Hsp70. BAG proteins bind through their BAG domain to the ATPase domain of Hsc70/Hsp70, and can modulate either positively or negatively the functions of the Hsc70/Hsp70 chaperone proteins. The BAG domain has been shown to contribute to the anti-apoptotic activity of BAG-family proteins. The anti-apoptotic activities of BAG-family proteins may be dependent on their interactions with Hsc70/Asp70 and/or binding to Bcl-2. In addition to the conserved BAG domain, BAG-family proteins also contain additional domains which enable them to interact with specific target proteins or to target them to specific locations within cells. The BAG family contains at least six family members, including BAG-1 and its various isoforms [including BAG-1S , BAG-1M (RAP46/HAP46), and BAG-1L, BAG2, BAG3 (CAIR-1; Bis,), BAG4 (SODD), BAG5 and BAG6 (Scythe, BAT3). The following amino acids (aa) lengths and molecular weights (kDa) have been described for human BAG proteins (reviewed in Takayama et al, 2001 and Doong et al, 2002): BAG-1 (230 aa., 34 kDa), BAG-1S (219 aa, 29 kDa), BAG-1M (274 aa, 46 kDa), BAG-1L (345 aa, 52 kDa), BAG-2 [212 aa; 24 kDa (Arndt et al. 2005)], BAG-3 (575 aa, 74 kDa), BAG-4 (456 aa; 60 kDa), BAG-5 ([442 aa; 51 kDa (Kalia et al. 2004)], and BAG-6 (1129 aa; 150 kDa).

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: BAG5