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HSP20/HSPB6 Overexpression Lysate

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Western Blot: Hsp20 Overexpression Lysate (Adult Normal) [NBL1-11759] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Hsp20.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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HSP20/HSPB6 Overexpression Lysate Summary

Description

HSP20/HSPB6 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC209637

Accession#: NM_144617

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
HSPB6

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
17 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 HSP20/HSPB6 Overexpression Lysate

  • FLJ32389
  • Heat shock 20 kDa-like protein p20
  • heat shock protein beta-6
  • heat shock protein, alpha-crystallin-related, B6
  • HSP20
  • HSPB6

Background

Hsp20 is a small heat shock protein related to Hsp25, Hsp27 and may form different hetercomplexes with these proteins. The specific physiological function of Hsp20 is not yet known. It is distributed ubiquitously in tissues, but is found in higher levels in skeletal, smooth and heart muscle. Under normal conditions, Hsp20 is diffusely distributed in the cytosol, but under heat stress conditions, it translocates to the nucleus. Unlike other heat shock proteins the amount of Hsp20 does not increase after heat shock. The Hsp20 was demonstrated to constitute up to 1.3% of the total cellular protein in vertebrate tissues, especially in muscle, and its expression is related to muscle contraction, specifically in slow-twitch muscle. Hsp20 may form different heterocomplexes with other Hsp's, such as alpha-crystalline and Hsp25. Phosphorylated form of Hsp20 is proposed to interact with monomeric actin whereas dephosphorylated form binds polymeric actin filaments. In normal conditions Hsp20 is diffusely disturbed in cytosol but under the heat stress it undergoes translocation to membrane fraction.

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 HSPB6