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Perforin Overexpression Lysate

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Western Blot: Perforin Overexpression Lysate (Adult Normal) [NBP2-10656] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Perforin.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Perforin Overexpression Lysate Summary

Description

Perforin Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC208940

Accession#: NM_001083116

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
PRF1

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
59.1 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 Perforin Overexpression Lysate

  • Cytolysin
  • FLH2
  • HPLH2
  • HPLH2lymphocyte pore forming protein
  • Lymphocyte pore-forming protein
  • MGC65093
  • P1
  • P1PFN1
  • perforin 1 (pore forming protein)
  • Perforin
  • perforin-1
  • PFP
  • PFPcytolysin
  • PRF1

Background

Perforin (PRF1) is a 70 kD cytolytic protein that is the major protein constituent of cytoplasmic granules of cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. Specifically, perforin enters the target cell's plasma membrand and creates a intercellular pore to mediate targeted cell lysis. PRF1 antibodies are useful tools for lysis research on both CTLs and NK cells.

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 Perforin Lysate (NBP2-10656) (0)

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Product General Protocols

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Video Protocols

WB Video Protocol

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Research Areas for Perforin Lysate (NBP2-10656)

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Blogs on Perforin.

You complete me: Natural killer cells need TGF-beta inhibition to effectively combat cancers
By Jamshed Arslan Pharm.D. Natural killer (NK) cells are lymphocytes of the innate immune system that were first discovered for their “natural” ability to kill cancer cells. To use NK cells as anti-cancer therapy, t...  Read full blog post.

Perforin a Protective Serial Killer
Secretory granule-mediated cell death is one of the the key mechanisms for elimination of virus-infected and transformed target cells by cytotoxic lymphocytes. Formation of the immunological synapse between an effector and a target cell leads to exocy...  Read full blog post.

Perforin Antibodies Reveal Links to Apoptosis and Immune Response
Perforin, also known as the pore-forming protein, pfp, is a 70 kD cytolytic protein expressed in the cytoplasmic granules of cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. It is one of the major effector molecules used by CTL and NK cel...  Read full blog post.

Perforin Antibodies for Detecting Immune System Diseases
 Perforin is a calcium-dependent pore forming cytolytic protein. Perforin is partially homologous to the terminal components of the membrane attack complex of complement and produces pores of up to 20nm in diameter on target membranes. Killer lymphocy...  Read full blog post.

Perforin Antibodies Reveal Links to Apoptosis and Immune Response
Perforin, also known as the pore-forming protein, pfp, is a 70 kD cytolytic protein expressed in the cytoplasmic granules of cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. It is one of the major effector molecules used by CTL and NK cel...  Read full blog post.

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Bioinformatics

Gene Symbol PRF1