hnRNP A1 Overexpression Lysate

Western Blot: hnRNP A1 Overexpression Lysate (Adult Normal) [NBP2-06143] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for hnRNP A1.

• Reactivity: Hu
• Applications: WB

hnRNP A1 Overexpression Lysate Summary

• Description:

  hnRNP A1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC211626
  
  Accession#: NM_031157
  
  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: HNRNPA1

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: 38.6 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 hnRNP A1 Overexpression Lysate

• Helix-destabilizing protein
• heterogeneous nuclear ribonucleoprotein A1
• heterogeneous nuclear ribonucleoprotein A1B protein
• heterogeneous nuclear ribonucleoprotein B2 protein
• heterogeneous nuclear ribonucleoprotein core protein A1
• hnRNP A1
• hnRNP core protein A1
• hnRNPA1
• hnRNP-A1
• HNRPA1MGC102835
• nuclear ribonucleoprotein particle A1 protein
• single-strand DNA-binding protein UP1
• Single-strand RNA-binding protein

Background

RNA polymerase II transcripts in the nucleus are in complex with several proteins called heterogeneous nuclear ribonucleoproteins (hnRNPs). These proteins are important in biological activities such as transcription, pre-mRNA processing, cytoplasmic mRNA translation and turnover. hnRNPs can be isolated either by immunoprecipitation or by sucrose gradient fractionation of cell extracts.1, 3, 4 Isolated hnRNPs consist of protein groups named A to U and many of these protein groups consist of more than one isoform. The major steady-state proteins of the isolated hnRNP complex are A1, A2, B1, B2, C1, and C2, with a range of molecular weight starting with 34 kDa up to 43 kDa.1, 3, 4 hnRNP-A1 is important in pre-mRNA processing and in mRNA export from the nucleus. The protein binds to its RNA target through a consensus RNA-binding site UAGGGU. The protein contains a 38-amino acid domain called M9, which is important for the interaction with the transportin protein, and therefore, for its import and export from the nucleus. RanGTP mediates dissociation of hnRNP-A1 from transportin. hnRNP-A1 is ubiquitously expressed with a higher expression in proliferating and/or transformed cells than in differentiated tissues.5

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: HNRNPA1