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ARF1 Overexpression Lysate

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Western Blot: ARF1 Overexpression Lysate (Adult Normal) [NBP2-09169] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ARF1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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ARF1 Overexpression Lysate Summary

Description

ARF1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC201240

Accession#: NM_001024227

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
ARF1

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
20.5 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 ARF1 Overexpression Lysate

  • ADP-ribosylation factor 1
  • ARF1

Background

The ADP-ribosylation factor (Arf) family comprises a group of structurally and functionally conserved 21 kDa proteins, which are members of the Ras superfamily of regulatory GTP-binding proteins. Arf is involved in intracellular protein traffic to and within the Golgi complex. Arf has a number of disparate activities including maintenance of organelle integrity, assembly of coat proteins, as a co-factor for cholera toxin and as an activator of phospholipase D. The Arf family is divided functionally into the Arf and the Arf-like (Arl) proteins. Arfs share more than 60% sequence identity, appear to be ubiquitous in eukaryotes, and are highly conserved evolutionarily.

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.

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Product General Protocols

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Research Areas for ARF1 Lysate (NBP2-09169)

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Blogs on ARF1.

Arf1: A New Focus In Cancer Drug Therapy
ARF1 (ADP-ribosylation factor 1) is a protein in the ARF gene family that is responsible for vesicular trafficking within the cell through its activation of phospholipase D. It is found in the cells golgi apparatus and its main function is intra-Golgi...  Read full blog post.

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Bioinformatics

Gene Symbol ARF1