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INSIG-2 Overexpression Lysate

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Western Blot: INSIG-2 Overexpression Lysate (Adult Normal) [NBL1-12004] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for INSIG-2.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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INSIG-2 Overexpression Lysate Summary

Description

INSIG-2 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC205274

Accession#: NM_016133

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
INSIG2

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
24.6 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 INSIG-2 Overexpression Lysate

  • INSIG2 membrane protein
  • INSIG-2
  • insulin induced gene 2
  • insulin induced protein 2
  • insulin-induced gene 2 protein
  • MGC26273

Background

The protein encoded by this gene is highly similar to the protein product encoded by gene INSIG1. Both INSIG1 protein and this protein are endoplasmic reticulum proteins that block the processing of sterol regulatory element binding proteins (SREBPs) by binding to SREBP cleavage-activating protein (SCAP), and thus prevent SCAP from escorting SREBPs to the Golgi. [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 INSIG-2 Lysate (NBL1-12004) (0)

There are no publications for INSIG-2 Lysate (NBL1-12004).
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Product General Protocols

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Video Protocols

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Research Areas for INSIG-2 Lysate (NBL1-12004)

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Blogs on INSIG-2.

SREBP: Gatekeeper of Cholesterol Homeostasis
SREBP1 (sterol-regulatory-element-binding protein 2) is a basic-helix-loop-helix-leucine zipper (bHLH-ZIP) transcription factor. It regulates sterol and cholesterol homeostasis by controlling enzymes involved in cholesterol synthesis and uptake, e.g. ...  Read full blog post.

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Bioinformatics

Gene Symbol INSIG2