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EAR2/NR2F6 Overexpression Lysate

Name: EAR2/NR2F6 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-13776
Price: 679 USD

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Western Blot: NR2F6 Overexpression Lysate (Adult Normal) [NBL1-13776] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NR2F6.

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Summary

Reactivity	HuSpecies Glossary
Applications	WB

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EAR2/NR2F6 Overexpression Lysate Summary

Description

EAR2/NR2F6 Transient Overexpression Lysate

Expression Host: HEK293T

Plasmid: RC201336

Accession#: NM_005234

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

NR2F6

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	42.8 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 EAR2/NR2F6 Overexpression Lysate

EAR2
EAR-2ERBA-related gene-2
EAR2nuclear receptor subfamily 2 group F member 6
ERBAL2
ERBAL2v-erb-a avian erythroblastic leukemia viral oncogene homolog-like 2
NR2F6
nuclear receptor subfamily 2, group F, member 6
V-erbA-related protein 2

Background

EAR2 is a nuclear orphan receptor related to the ERB-A superfamily and the COUP-TF family of transcription factors. EAR2 has been shown to be a transcriptional co-repressor of the lutenizing hormone receptor and the thyroid hormone receptor. It has also been shown to associate with negative transcription regulatory elements, and repress promoter activity. Alternate names for EAR2 include nuclear receptor subfamily 2 group F member 6, orphan nuclear receptor EAR-2, V-erbA-related protein, NR2F6, and ERBAL2.

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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EAR2/NR2F6
Overexpression Lysate

NBL1-13776

Species: Hu

Applications: WB

Publications for EAR2/NR2F6 Lysate (NBL1-13776) (0)

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Product General Protocols

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Loss of miR-101 promotes cancer metastasis through de-repression of COUP-TFII expression.(a) LNCaP and PC3 cells were individually treated with 50 nM of miR-101 and miR-27a mimic for 48 h and then invasion assays were performed for an additional 16 h (n=3). *P1.5-fold are represented as a heatmap. The gene symbol of FOXM1 and CENPF genes after knockdown of COUP-TFII are annotated in red colour. The others were FOXM1 and CENPF co-regulated genes annotated in black colour. siCtrl: knockdown scramble control; siCII: knockdown of COUP-TFII. (b) A representative western blot shows the level of COUP-TFII in PC3 cells transfected with two different siRNAs against COUP-TFII for 72 h (upper panel). Levels of FOXM1 and CENPF are quantified by RT-qPCR (lower panel) (n=3). *P<0.05 (two-sided Student's t-test) compared with the control group (siCON). (c) A cartoon shows the FOXM1 and CENPF loci. COUP-TFII-binding sites are annotated by a black rectangle (upper panel). ChIP results are shown in the PC3 cells treated with control and two different siRNAs against COUP-TFII for 72 h (lower panel) (n=3). *P<0.05 (two-sided Student's t-test) compared with control. (d) FOXM1 and CENPF promoter activities were measured in PC3 cells carrying an inducible COUP-TFII gene in the absence or the presence of doxycycline for 48 h (n=3). *P<0.05 (two-sided Student's t-test). (e) FOXM1 and CENPF promoter activities were measured in control or stably knocked-down COUP-TFII in PC3 cells for 48 h (n=3). *P<0.05 (two-sided Student's t-test). (f) PC3 cells carrying with inducible COUP-TFII-Flag gene were treated with or without doxycycline for 48 h and then transfected with individual siRNA against FOXM1 and CENPF or double knockdown of FOXM1 and CENPF for an additional 48 h. western blot (left panel) shows the level of COUP-TFII, CENPF and FOXM1 expression. Invasion assay (right panel) was performed by using those cells (n=3). *P<0.05 (two-sided Student's t-test) compared with control. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/ncomms11418), licensed under a CC-BY license. Not internally tested by R&D Systems.

COUP-TF II/NR2F2 Antibody (H7147) [Unconjugated]
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Bioinformatics

Gene Symbol

NR2F6

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

EAR2/NR2F6 Overexpression Lysate

Species Glossary

Images

Product Details

Order Details

EAR2/NR2F6 Overexpression Lysate Summary

EAR2/NR2F6 Transient Overexpression Lysate

Applications/Dilutions

Packaging, Storage & Formulations

Lysate Details for Array

Notes

Alternate Names for EAR2/NR2F6 Overexpression Lysate

Background

Limitations

Customers Who Viewed This Item Also Viewed...

Publications for EAR2/NR2F6 Lysate (NBL1-13776) (0)

Reviews for EAR2/NR2F6 Lysate (NBL1-13776) (0)

Product General Protocols

Video Protocols

FAQs for EAR2/NR2F6 Lysate (NBL1-13776) (0)

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Bioinformatics