ARFIP1 Overexpression Lysate

Name: ARFIP1 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-07658

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-07658

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for ARFIP1 Overexpression Lysate

ARFIP1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC211267

Accession#: NM_001025593

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.

Product Specifications

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.

TMW

38.4 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.

Type

Overexpression

Scientific Data Images for ARFIP1 Overexpression Lysate

Western Blot: ARFIP1 Overexpression Lysate [NBL1-07658]

Western Blot: ARFIP1 Overexpression Lysate (Adult Normal) [NBL1-07658] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ARFIP1.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: ARFIP1

ADP-ribosylation factors, or ARFs, enhance the ADP ribosyltransferase activity of cholera toxin and are implicated in vesicle transport between endoplasmic reticulum and the Golgi complex. Arfaptin 1 is recruited from the cytosol to Golgi membranes by ARFs in a guanosine 5'-O-(3-thiotriphosphate)-dependent and brefeldin A-sensitive manner but is not a constituent of coatomer. Arfaptin 1 binds to nonmyristoylated GTP-bound ARF3, but not to GDP-bound ARF3, and also to ARF1, another class I ARF. It binds with lower affinity to ARF5, a class II ARF, and with very little affinity to ARF6, a class III ARF. POR1 (also designated arfaptin 2) was first isolated as a Rac 1 binding protein necessary for Rac mediated Actin polymerization and the subsequent formation of membrane ruffles and lamellipodia. POR1 has also been shown to interact with the ADP ribosylation factor ARF6, a GTPase that associates with the plasma membrane and intracellular endosome vesicles, in a GTP dependent manner. The association of POR1 with ARF6 stimulates induction of Actin polymerization. POR1 appears to play a regulatory role through multiple signaling pathways in the reorganization of the cytoskeletal structure.

Alternate Names

ADP-ribosylation factor interacting protein 1, ADP-ribosylation factor-interacting protein 1, arfaptin 1, arfaptin-1, HSU52521, MGC117369

Gene Symbol

ARFIP1

Additional ARFIP1 Products

Product Documents for ARFIP1 Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for ARFIP1 Overexpression Lysate

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.

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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