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SPARC Overexpression Lysate

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Western Blot: SPARC Overexpression Lysate (Adult Normal) [NBL1-16372] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for SPARC.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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SPARC Overexpression Lysate Summary

Description

SPARC Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC209964

Accession#: NM_003118

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
SPARC

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
34.5 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 SPARC Overexpression Lysate

  • Basement-membrane protein 40
  • BM-40
  • ONcysteine-rich protein
  • Osteonectin
  • Secreted protein acidic and rich in cysteine
  • secreted protein, acidic, cysteine-rich (osteonectin)
  • SPARC

Background

Appears to regulate cell growth through interactions with the extracellular matrix and cytokines. Binds calcium and copper, several types of collagen, albumin, thrombospondin, PDGF and cell membranes. There are two calcium binding sites; an acidic domain that binds 5 to 8 Ca(2+) with a low affinity and an EF-hand loop that binds a Ca(2+) ion with a high affinity.

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 SPARC Lysate (NBL1-16372) (0)

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Product General Protocols

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Video Protocols

WB Video Protocol

FAQs for SPARC Lysate (NBL1-16372). (Showing 1 - 1 of 1 FAQ).

  1. I would like to clarify some of the questions including is there any possibility to use this product to overexpress SPARC in C9 rat liver cell lines or other cell lines.
    • Our SPARC over-expression lysate with catalogue number NBL1-16372 was created in HEK293 cells and is intended for use as a positive control when Western blotting. The HEK293 cells were transfected with a plasmid based on accession number NM_003118, and you would receive 100ug of the over-expression lysate plus 100ug of a control (HEK293 transfected with the empty vector).

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Research Areas for SPARC Lysate (NBL1-16372)

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Blogs on SPARC.

Caspase 8 - a key mediator of apoptosis
Programmed cell death via apoptosis is a key controlled physiological process instigated by the cell death receptor family, their ligands, and the caspase cysteine protease family. All caspases exist in a precursor form that contains a prodomain, a...  Read full blog post.

GAPDH: More than a housekeeping gene
GAPDH is a 146kD tetramer glycolytic pathway metabolic enzyme composed of four 30-40 kDa subunits. It is responsible for reversibly phosphorylating its substrate glyceraldehyde 3-phosphate within the glycolytic pathway.  Apart from its role in gl...  Read full blog post.

GAPDH (Glyceraldehyde 3-Phosphate Dehydrogenase)
GAPDH is a 146 kD tetramer glycolytic pathway metabolic enzyme responsible for reversibly phosphorylating glyceraldehyde 3-phosphate. It may have other possible functions in transcriptional activation. GAPDH is highly expressed due to this housekeepin...  Read full blog post.

FLICE, FLICE, baby
Cell death via apoptosis is a fundamental cellular function triggered by the cell death receptor family and their ligands which signal through downstream adaptor molecules and the caspase protease family. All caspases exist in a precursor form compose...  Read full blog post.

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Bioinformatics

Gene Symbol SPARC