DCC Overexpression Lysate

Name: DCC Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-07775

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-07775

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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 DCC Overexpression Lysate

DCC Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC218506

Accession#: NM_005215

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

158.3 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.

Type

Overexpression

Scientific Data Images for DCC Overexpression Lysate

Western Blot: DCC Overexpression Lysate [NBP2-07775]

Western Blot: DCC Overexpression Lysate (Adult Normal) [NBP2-07775] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for DCC.

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: DCC

Invasive, metastatic colon cancer arises from pre-existing adenomas in both familial and sporadic cases and is characterized by the presence of multiple chromosomal alterations. In the progression from intramucosal carcinoma to invasive carcinoma, allelic loss on the long arm of chromosome 18 is frequently observed. Recently, a gene on 18q21.3 termed DCC, for deleted in colorectal carcinoma, has been isolated and shown to be deleted or mutated in both sporadic and inherited colon carcinoma whereas normal colonic tissue retained and expressed the gene. Furthermore, loss of DCC expression appears to accompany progression of adenomas to metastatic carcinoma. The inactivation of DCC suggests that it is a tumor suppressor gene. A functional role for DCC as a tumor suppressor gene is indicated by experiments demonstrating that introduction of a normal chromosome 18 into the human colon tumor cell lines COKFu or SW480.7 lacking DCC suppresses tumorigenicity. Analysis of the amino acid sequence deduced from cDNA clones indicates that DCC is a transmembrane glycoprotein consisting of 1447 amino acids with an extracellular domain having extensive similarity to neural cell adhesion molecules (NCAM), a single transmembrane segment, and a unique cytoplasmic domain. The high degree of similarity to NCAMs suggests that DCC is involved in cell-cell interactions essential to the differentiated state of the colonic epithelium. DCC expression has been observed in all tissues examined except liver with the highest level of expression in brain tissue. Alterations in DCC expression have also been observed in pancreatic adenocarcinoma and esophogeal carcinoma. However, DCC mRNA is expressed at very low levels, requiring reverse transcription followed by PCR amplification for unambiguous detection.

Long Name

Deleted in Colorectal Cancer

Alternate Names

Colorectal cancer suppressor, CRC18, CRCR1, deleted in colorectal cancer protein, deleted in colorectal carcinoma, IGDCC1colorectal tumor suppressor, Immunoglobulin superfamily DCC subclass member 1, immunoglobulin superfamily, DCC subclass, member 1, netrin receptor DCC, Tumor suppressor protein DCC

Gene Symbol

DCC

Additional DCC Products

Product Documents for DCC Overexpression Lysate

Product Datasheets

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COA

SDS

Download SDS

Product Specific Notices for DCC 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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