PCNA Overexpression Lysate

Images

 
Western Blot: PCNA Overexpression Lysate (Adult Normal) [NBL1-14184] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PCNA.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

PCNA Overexpression Lysate Summary

Description

PCNA Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC201741

Accession#: NM_002592

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
PCNA

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot.

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
28.6 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.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

Alternate Names for PCNA Overexpression Lysate

  • cyclin
  • DNA polymerase delta auxiliary protein
  • MGC8367
  • PCNA
  • proliferating cell nuclear antigen

Background

The protein encoded by this gene is found in the nucleus and is a cofactor of DNA polymerase delta. The encoded protein acts as a homotrimer and helps increase the processivity of leading strand synthesis during DNA replication. In response to DNA damage, this protein is ubiquitinated and is involved in the RAD6-dependent DNA repair pathway. Two transcript variants encoding the same protein have been found for this gene. Pseudogenes of this gene have been described on chromosome 4 and on the X chromosome. [provided by RefSeq]. Transcript Variant: This variant (1) represents the longer transcript. Variants 1 and 2 encode the same protein. 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 PCNA Lysate (NBL1-14184) (0)

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Product General Protocols

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Video Protocols

WB Video Protocol

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PCNA (Proliferating cell nuclear antigen, polymerase delta auxiliary protein)
PCNA is a nuclear protein essential for DNA replication as well as DNA excision and mismatch repair pathways. It coordinates the recruitment and association of needed components during both of these processes, both of which are essential for cell cycl...  Read full blog post.

PCNA Antibodies: Marking Cell Proliferation & DNA Replication
Proliferating Cell Nuclear Antigen (PCNA), also known as the polymerase delta auxiliary protein, is a nuclear protein essential for DNA replication as well as DNA excision and mismatch repair pathways. It has a large role in cell cycle regulation and ...  Read full blog post.

PCNA is a Universal Marker of Proliferating Cells
Proliferating cell nuclear antigen (PCNA) is an evolutionarily well-conserved protein found in all eukaryotic species as well as in Archaea. PCNA was first shown to be involved in DNA replication. However PCNA functions are associated with other vital...  Read full blog post.

Using PCNA as an Antibody Marker
PCNA antibodies are useful biomarkers in DNA repair studies. PCNA is one of several proteins essential for the completion of nucleotide excision repair, a multi-stage process involving 20 - 30 proteins, and an important factor in repairing damage and ...  Read full blog post.

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Bioinformatics

Gene Symbol PCNA