Prominin 2 Overexpression Lysate

Western Blot: Prominin 2 Overexpression Lysate (Adult Normal) [NBL1-14811] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Prominin 2.

• Reactivity: Hu
• Applications: WB

Prominin 2 Overexpression Lysate Summary

• Description:

  Prominin 2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC211605
  
  Accession#: NM_144707
  
  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: PROM2

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: 91.7 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 Prominin 2 Overexpression Lysate

• hPROML2
• MGC138714
• PROM2
• PROM-2
• Prominin 2
• prominin-2
• Prominin-like protein 2
• prominin-related protein
• PROML2

Background

Prominin 2 is a large-scale effort, termed the Secreted Protein Discovery Initiative (SPDI), was undertaken to identify novel secreted and transmembrane proteins. Several criteria were applied to identify and characterize these new and novel genes. First, a biological signal sequence trap in yeast cells was utilized to identify cDNA clones encoding putative secreted proteins, second strategy utilized various algorithms that recognize features such as the hydrophobic properties of signal sequences to identify putative proteins encoded by expressed sequence tags (ESTs) from human cDNA libraries and finally, ESTs for protein sequence similarity to a set of known receptors and their ligands with the BLAST algorithm. Based on these criteria isolation of full length cDNA clones for each of these genes resulted in identification of more than 1000 novel proteins. One of these new genes is Prominin2. Prominin 2 is a 112 kDa glycoporotein structurally related to Prominin 1 (CD133) although amino acid similarity is not more than 30% but their genomic organization is strikingly similar (1). Like Prominin1, the prominin 2 exhibit similar membrane topology with 5 trans-membrane domains and two large glycosylated extracellular domains. Similar to Prominin1 localization, the Prominin 2 is also associated with membrane protrusions of the epithelial cells from adult kidney, and all along the digestive track and other epithelial tissues. One striking difference between Prominin1 and Prominin 2 expression lies in its conspicuous absence in eye tissue (2). Prominin 2, along with other genes (AP1M2, MAL2, PRSS8 and FLJ20171) expression is differentially expressed in chromophobe renal cell carcinoma compared to benign oncocytoma consistently and this marker can be used to differentiate RCC form benign oncocytoma (3). Prominin 2 is an androgen-responsive genes proapoptotic membrane protein that is expressed in human prostate and human prostate cancer cell lines with highest levels in less aggressive LNCaP cell and low expression in highly aggressive PC3 and DU145 cells suggesting a role of Prominin 2 in down-regulation of aggressiveness of prostate cancer cells (4).

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: PROM2