NHERF-1 Overexpression Lysate

Western Blot: NHERF-1 Overexpression Lysate (Adult Normal) [NBL1-16198] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NHERF-1.

• Reactivity: Hu
• Applications: WB

NHERF-1 Overexpression Lysate Summary

• Description:

  NHERF-1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC201653
  
  Accession#: NM_004252
  
  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: NHERF1

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: 38.7 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 NHERF-1 Overexpression Lysate

• EBP50NHERF-1
• ezrin-radixin-moesin binding phosphoprotein-50
• Ezrin-radixin-moesin-binding phosphoprotein 50
• Na(+)/H(+) exchange regulatory cofactor NHE-RF1
• Na+/H+ exchange regulatory co-factor
• NHERF1solute carrier family 9 (sodium/hydrogen exchanger), isoform 3 regulator 1
• NHERFNPHLOP2
• Regulatory cofactor of Na(+)/H(+) exchanger
• sodium/hydrogen exchanger regulatory factor 1
• Sodium-hydrogen exchanger regulatory factor 1
• solute carrier family 9 (sodium/hydrogen exchanger), isoform 3 regulatoryfactor 1
• solute carrier family 9 (sodium/hydrogen exchanger), member 3 regulator 1
• Solute carrier family 9 isoform A3 regulatory factor 1

Background

Members of the ezrin (VIL2; MIM 123900)-radixin (RDX; MIM 179410)-moesin (MSN; MIM 309845) (ERM) protein family are highly concentrated in the apical aspect of polarized epithelial cells. These cells are studded with microvilli containing bundles of actin filaments, which must attach to the membrane to assemble and maintain the microvilli. The ERM proteins, together with merlin, the NF2 (MIM 607379) gene product, are thought to be linkers between integral membrane and cytoskeletal proteins, and they bind directly to actin in vitro. Actin cytoskeleton reorganization requires the activation of a sodium/hydrogen exchanger (SLC9A3; MIM 182307). SLC9A3R1 is an ERM-binding protein.[supplied by OMIM]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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: NHERF1