AGO1/EIF2C1 Overexpression Lysate

Western Blot: AGO1/EIF2C1 Overexpression Lysate (Adult Normal) [NBL1-10182] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for AGO1/EIF2C1.

• Reactivity: Hu
• Applications: WB

AGO1/EIF2C1 Overexpression Lysate Summary

• Description:

  AGO1/EIF2C1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC209163
  
  Accession#: NM_012199
  
  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: AGO1

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: 97 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 AGO1/EIF2C1 Overexpression Lysate

• AGO1Q99
• argonaute 1
• argonaute1
• DKFZp686M13167
• eIF2C 1
• eIF-2C 1
• EIF2C
• Eukaryotic translation initiation factor 2C 1
• eukaryotic translation initiation factor 2C, 1
• GERP95
• Golgi Endoplasmic Reticulum protein 95 kDa
• hAgo1
• protein argonaute-1
• Putative RNA-binding protein Q99

Background

Required for RNA-mediated gene silencing (RNAi). Binds to short RNAs such as microRNAs (miRNAs) or short interfering RNAs (siRNAs), and represses the translation of mRNAs which are complementary to them. Lacks endonuclease activity and does not appear to cleave target mRNAs. Also required for transcriptional gene silencing (TGS) of promoter regions which are complementary to bound short antigene RNAs (agRNAs). Interacts with DDB1, DDX5, DDX6, DHX30, DHX36, DDX47, DICER1, EIF2C2/AGO2, ELAVL1, HNRNPF, IGF2BP1, ILF3, IMP8, MATR3, MOV10, PABPC1, PRMT5, RBM4, SART3, TNRC6B, UPF1 and YBX1. Associates with polysomes and messenger ribonucleoproteins (mNRPs). Interacts with LIMD1, WTIP and AJUBA.

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.

FAQs for AGO1/EIF2C1 Lysate (NBL1-10182). (Showing 1 - 1 of 1 FAQ).

1. I am interested in possibly ordering the AGO1 lysate as a positive control for my western blot. Can you please tell me the quantity to add per lane of western blot?
   • We recommend adding 10ug per lane of this lysate for Western blot.

Bioinformatics

• Gene Symbol: AGO1