IGFBP-6 Overexpression Lysate

Western Blot: IGFBP6 Overexpression Lysate (Adult Normal) [NBL1-11876] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for IGFBP6.

• Reactivity: Hu
• Applications: WB

IGFBP-6 Overexpression Lysate Summary

• Description:

  IGFBP-6 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC204060
  
  Accession#: NM_002178
  
  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: IGFBP6

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: 25.1 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 IGFBP-6 Overexpression Lysate

• IBP6
• IBP-6
• IGF binding protein 6
• IGF-binding protein 6
• IGFBP6
• IGFBP-6
• insulin-like growth factor binding protein 6
• insulin-like growth factor-binding protein 6

Background

Data show that an increase in JNK activation in the presence of NFkappaB inhibition significantly increased the expression of IGFBP6. These studies provide evidence that over expression of IGFBP-6 suppresses human and murine osteoblast differentiation, that IGFBP-6 and LMP-1 physically interact, and supports the conclusion that this interaction may be functionally relevant. IGFBP-6 is the effector of tumor suppressor activity of SEMA3B. Observational study of gene-disease association. (HuGE Navigator) IGFBP-6 is translocated to the nucleus with functional consequences, and different members of the IGFBP family have specific nuclear import mechanisms. Toxic effect of TCDD on osteogenesis through altering IGFBP6 gene expression in osteoblasts is reported. p38 MAPK is involved in IGFBP-6-induced IGF-independent rhabdomyosarcoma cell migration. determination of blood levels in adult patients with severe liver disease before and after orthotopic liver transplantation. Inhibition of human osteoblast marker gene expression by retinoids is mediated in part by insulin-like growth factor binding protein-6. A close correlation exists between residues of the carboxyl-terminal domain of IGFBP6 undergoing conformational change in 15N NMR spectroscopy studies with those that disappeared or broadened upon insulin-like growth factor (IGF-II) binding. C-domain of IGFBP-6 consists of a thyroglobulin type 1 fold comprising an alpha-helix followed by a loop, a three-stranded antiparallel beta-sheet incorporating a second loop, and a disulfide-bonded flexible third loop. The expression of certain IGFBP is significantly altered in renal cell carcinoma. CCI-779 acts additively with IGFBP-6 to reduce rhabdomyosarcoma growth both in vitro and in vivo. may function as an antiproliferative molecule suppressing mitogenic effects of insulin-like growth factors on Malassez cells. The IGFBP-6 protein is synthesized as propeptides with a hydrophobic leader sequence, removal of which yields a mature protein composed of 3 recognizable domains of similar size. transgenic human IGFBP-6 mice may be considered a new tool for studies of the involvement of the brain IGF system in metabolism control and obesity.

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: IGFBP6