We tested both NBP1-49669 and R-079-100 by WB, but failed to obtain signal. May we ask your suggestion for positive control? We found NBL1-17341 may be an option for R-079-100, but is there any other lysate which can be suggested and is easier to get?

Name: Tight Junction Protein 1 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-10877

In our catalog, NBP2-10877 as well as NBP2-05057 are the products that are suggested as lysates for ZO-1 tight junction protein antibody (NBP1-49669). You already know NBL1-17341 may be an option for VRL1 Antibody (R-079-100). I do not see any other products in our collection that will go along ZO-1 tight junction protein antibody (NBP1-49669) and VRL1 antibody (R-079-100).

Tight Junction Protein 1 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-10877

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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 Tight Junction Protein 1 Overexpression Lysate

Tight Junction Protein 1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC222836

Accession#: NM_003257

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

195.3 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 Tight Junction Protein 1 Overexpression Lysate

Western Blot: Tight Junction Protein 1 Overexpression Lysate [NBP2-10877]

Western Blot: Tight Junction Protein 1 Overexpression Lysate (Native) [NBP2-10877] - Tight Junction Protein 1 Overexpression Lysate (Native) Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ZO-1 tight junction protein.

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: Tight Junction Protein 1

Members of this family are involved in epithelial and endothelial intercellular junctions. They each contain at least one PSD95/Dlg/ZO-1 (PDZ) domain, a Src homology 3 (SH3) domain, and an enzymatically inactive guanylate kinase domain. PDZ domains are 90-amino acid protein-protein binding domains that recognize at least a 3-residue peptide motif in the COOH termini of their binding partners. PDZ domain-containing proteins, like ZO-1, typically act as scaffolding proteins that organize membrane receptors and cytosolic proteins into multimeric signaling complexes often at the sites of cell-cell contact. The effectiveness and stability of the epithelial barrier depends on a complex of proteins composing different intercellular junctions, which include tight junctions, adherens junctions, and desmosomes. ZO-1 is a peripheral membrane protein bound on the cytoplasmic surface of junctional contacts and is expressed in all tight junctions regardless of their properties. ZO-1 immunoprecipitates with its family member ZO-2. ZO-1 was shown to undergo tyrosine phosphorylation during tight junction formation and remodeling. Two different isoforms of ZO-1, alpha-minus and alpha-plus, have been described, which result from alternative splicing of an mRNA encoded by a single gene. The ZO-1 alpha-plus contains an 80 amino acids motif called alpha which is not present in ZO-1 alpha-minus. The alpha-containing isoform is found in most epithelial cell junctions. The short isoform (ZO-1 alpha-minus) is found both in endothelial cells and the highly specialized epithelial junctions of renal glomeruli and Sertoli cells of the seminiferous tubules. This difference in distribution provides molecular distinction among tight junctions.

Alternate Names

DKFZp686M05161, MGC133289, Tight junction protein 1, tight junction protein 1 (zona occludens 1), tight junction protein ZO-1, TJP1, ZO1, ZO-1, zona occludens 1, Zona occludens protein 1, zonula occludens 1 protein, Zonula occludens protein 1

Gene Symbol

TJP1

Additional Tight Junction Protein 1 Products

Product Documents for Tight Junction Protein 1 Overexpression Lysate

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COA

SDS

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Product Specific Notices for Tight Junction Protein 1 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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