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ARF4 Overexpression Lysate

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Western Blot: ARF4 Overexpression Lysate (Adult Normal) [NBL1-07652] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ARF4.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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ARF4 Overexpression Lysate Summary

Description

ARF4 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC204984

Accession#: NM_001660

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
ARF4

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
20.3 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 ARF4 Overexpression Lysate

  • ADP-ribosylation factor 4
  • ARF2ADP-ribosylation factor 2

Background

This gene is a member of the human ARF gene family whose members encode small guanine nucleotide-binding proteins that stimulate the ADP-ribosyltransferase activity of cholera toxin and play a role in vesicular trafficking and as activators of phospholipase D. The gene products include 5 ARF proteins and 11 ARF-like proteins and constitute one family of the RAS superfamily. The ARF proteins are categorized as class I, class II and class III; this gene is a class II member. The members of each class share a common gene organization. The ARF4 gene spans approximately 12kb and contains six exons and five introns. This gene is the most divergent member of the human ARFs. Conflicting map positions at 3p14 or 3p21 have been reported for this gene. [provided by RefSeq]. 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.

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Publications for ARF4 Lysate (NBL1-07652) (0)

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Product General Protocols

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Video Protocols

WB Video Protocol

FAQs for ARF4 Lysate (NBL1-07652). (Showing 1 - 1 of 1 FAQ).

  1. I was looking for antibodies against human ARF4 and ARF5 and I need to be able to distinguish between the two proteins. Do you have antibodies specific for each that don't cross react with the other protein?
    • The sequence similarity between ARF4 and ARF5 shows very high homology between the two proteins. We have not specifically tested for cross-reactivity between the antibodies, but I think it's very likely that they will cross react.

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Bioinformatics

Gene Symbol ARF4