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ROBO4 Overexpression Lysate

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Western Blot: Robo4 Overexpression Lysate (Adult Normal) [NBL1-15473] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Robo4.
Western Blot: Robo4 Overexpression Lysate (Adult Normal) [NBL1-15473] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Robo4.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

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ROBO4 Overexpression Lysate Summary

Description

ROBO4 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC213730

Accession#: NM_019055

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
ROBO4

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 (cat# 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
107.3 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 ROBO4 Overexpression Lysate

  • ECSM4
  • FLJ20798
  • Magic roundabout
  • MGC133352
  • MGC133353
  • MRB
  • ROBO4
  • roundabout homolog 4
  • roundabout homolog 4, magic roundabout (Drosophila)

Background

Roundabouts (ROBO) are cell-surface receptors that mediate repulsive signaling mechanisms at the central nervous system midline. However, ROBOs may also mediate attraction mechanisms in the context of vascular development. ROBO4 is a novel roundabout protein which is restricted in expression to endothelial cells in vitro and sites of angiogenesis in vivo. Because it is not expressed on normal endothelial cells in vivo, it is a promising tumor endothelial marker.

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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Bioinformatics

Gene Symbol ROBO4