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Nanog Overexpression Lysate

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Western Blot: Nanog Overexpression Lysate (Adult Normal) [NBL1-13465] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Nanog.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Nanog Overexpression Lysate Summary

Description

Nanog Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC210243

Accession#: NM_024865

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
NANOG

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
34.4 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 Nanog Overexpression Lysate

  • FLJ12581
  • FLJ40451
  • hNanog
  • homeobox protein NANOG
  • Homeobox transcription factor Nanog
  • homeobox transcription factor Nanog-delta 48
  • Nanog homeobox
  • Nanog

Background

Nanog is one of the molecular markers suitable for recognizing the undifferentiated state of stem cells in the mouse and human. Nanog is a divergent homeodomain protein that directs propagation of undifferentiated ES cells. Nanog mRNA is present in pluripotent mouse and human cell lines, and absent from differentiated cells. Nanog-deficient ES cells lose pluripotency and differentiate into extraembryonic endoderm lineage.

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 Nanog Lysate (NBL1-13465) (0)

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Product General Protocols

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Video Protocols

WB Video Protocol

FAQs for Nanog Lysate (NBL1-13465) (0)

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Research Areas for Nanog Lysate (NBL1-13465)

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Blogs on Nanog.

Breast cancer stem cells survive chemotherapy through S100A10-ANXA2-SPT6 interaction that epigenetically promotes OCT4-mediated stemness
By Jamshed Arslan, Pharm D, PhDBreast cancer is the most common cancer among women that causes the greatest number of cancer-related deaths worldwide. After radiotherapy or cytotoxic chemotherapy like paclitax...  Read full blog post.

Deriving neural precursor cells from human induced pluripotent stem cells
By Jennifer Sokolowski, MD, PhD.Human induced pluripotent stem cells (iPSCs) can be used to create models of human organ systems and are useful for a) ascertaining the mechanisms underlying pathological conditions...  Read full blog post.

Stemness for Surviving Hypoxia: TGF-beta/Smad Signaling in Multiple Myeloma
By Jamshed Arslan Pharm.D. Multiple myeloma (MM) is a cancer of antibody-producing plasma cells. The bone marrow (BM) of MM patients is hypoxic, and MM cells overexpress many cancerous genes that are regulated by hy...  Read full blog post.

KLF4 as a transcription factor in stem cell differentiation
Kru¨ppel-like factors (KLFs) are evolutionarily conserved zinc finger transcription factors that play a role in cell differentiation, proliferation, and pluripotency. KLF4 has specifically been tied to many diverse cellular processes, including sel...  Read full blog post.

Nanog is a Master Controller of ES cell Pluripotency
Nanog, a homeodomain (HD) transcription factor, plays a critical role in the maintenance of embryonic stem (ES) cell self-renewal. Transcription regulator involved in inner cell mass and ES cell proliferation and self-renewal. Imposes pluripotency on ...  Read full blog post.

Sox2 and Oct4: Roles in Embryonic Stem Cell Pluripotency
Embryonic stem (ES) cells are cells derived from the inner cell mass of the blastocyst, an early-stage embryo. ES cells are distinguished from other cells due to their pluripotency, which is the ability to differentiate into any different type of cell...  Read full blog post.

An Unlikely Pairing: The SOX2 Antibody and Breast Cancer
SOX2 encodes a member of the SOX (SRY-related HMG-box) family of transcription factors, which play a vital role in embryonic development. SOX2 antibody research has identified Sox2 as a key transcription factor in pluripotent stem cells. We at Novus B...  Read full blog post.

Fluorescence Activated Cell Sorting Antibody Techniques
Recently, we at Novus Biologicals added several embryonic stem cell marker products to our antibody catalog, validated for use in fluorescent activated cell sorting (FACS) assays. They included Cripto1, PODXL, SSEA, OCT4, Nanog, SOX2, TRA-1, TERT and ...  Read full blog post.

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Bioinformatics

Gene Symbol NANOG