MMP20 Overexpression Lysate

Western Blot: MMP20 Overexpression Lysate (Adult Normal) [NBL1-13156] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for MMP20.

• Reactivity: Hu
• Applications: WB

MMP20 Overexpression Lysate Summary

• Description:

  MMP20 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC220786
  
  Accession#: NM_004771
  
  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: MMP20

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: 52 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 MMP20 Overexpression Lysate

• AI2A2
• EC 3.4.24
• EC 3.4.24.-
• EC 3.4.24.22
• EC 3.4.24.35
• Enamel metalloproteinase
• Enamelysin
• matrix metallopeptidase 20
• matrix metalloproteinase 20 (enamelysin)
• matrix metalloproteinase-20
• MMP20
• MMP-20

Background

MMP-20 (enamelysin) is a recently discovered member of the MMP family. This enzyme is involved in the degradation of the various components of the extracellular matrix (ECM) during development, haemostasis and pathological conditions. MMP-20 expression has been found to be restricted to the enamel organ and due to its unique expression pattern, may primarily be involved in the turnover of these molecules during tooth development. The enamel matrix proteins amelogenin, ameloblastin, and enamelin are expressed during the developmental time period, suggesting that MMP-20 may play a role in their hydrolysis (1). Recombinant human enamelysin is able to degrade amelogenin, the major protein component of the enamel matrix. On the basis of its degrading activity on amelogenin, and its highly restricted expression to dental tissues, we suggest that human enamelysin plays a central role in the process of tooth enamel formation (2). Enamelysin null mice do not process amelogenin properly, possesses an altered enamel matrix and rod pattern, has hypoplastic enamel that delaminates from the dentin, and has a deteriorating enamel organ morphology as development progresses (3).

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: MMP20