mCerulean3-Bcl-2-s2193 | Addgene 166750 product information

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product summary

company name :

Addgene

product type :

cDNA

product name :

mCerulean3-Bcl-2-s2193

catalog :

166750

citations: 2

Reference
Osterlund E, Hirmiz N, Pemberton J, Nougarède A, Liu Q, Leber B, et al. Efficacy and specificity of inhibitors of BCL-2 family protein interactions assessed by affinity measurements in live cells. Sci Adv. 2022;8:eabm7375 pubmed publisher
Liu Q, Osterlund E, Chi X, Pogmore J, Leber B, Andrews D. Bim escapes displacement by BH3-mimetic anti-cancer drugs by double-bolt locking both Bcl-XL and Bcl-2. elife. 2019;8: pubmed publisher

product information

Catalog Number :

166750

Product Name :

mCerulean3-Bcl-2-s2193

article :

doi	10.7554/eLife.37689
id	28216034
pubmed_id	30860026

bacterial resistance :

Blasticidin

cloning :

backbone

s2193

backbone_mutation

backbone_origin

backbone_size

promoter

sequencing_primer_3

sequencing_primer_5

vector_types

Mammalian Expression

growth notes :

Transfection of this plasmid will express mCerulean3-fused to the N-terminus of Bcl-2. There is a flexible, 7 amino acid linker between SGLRSTR , between mCerulean3 and Bcl-2. Selection with Blasticidin was used to generate BMK-DKO cells stably expressing this plasmid. Expression under control of Human ferritin promoter. Glysine 237 of Bcl-2 is mutated to Serine, with no effect on anti-apoptotic function. Please note coding region of Bcl-2 is from human DNA described in 1994 (Janiak et al., 1994) published in JBC. There is a M157I mutation in this Bcl-2 sequence. This mutation was in the original clone and has been used to inhibit apoptosis by our group and many others around the world since that time. It has always been our assumption that this mutation is a naturally occurring variant. Our clone has been used in many publications as the plasmid was widely distributed when it was first assembled and published. Janiak F, Leber B, Andrews DW. Assembly of Bcl-2 into microsomal and outer mitochondrial membranes. J Biol Chem. 1994 Apr 1;269(13):9842-9. PMID: 8144576.

growth strain :

Express mCerulean3 fused to the N-terminus of the Bcl-2 family protein, Bcl-2 in a plasmid with Blasticidin resistance. Used to make stable BMK-DKO cell lines.

origin :

pi :

alt_names

cloning

clone_method	Restriction Enzyme
cloning_site_3	Unknown
cloning_site_5	Unknown
promoter	Human ferritin
sequencing_primer_3	TTACCCCTCTAGACCTGGAAAG
sequencing_primer_5	CCTTGAGTTTTGAGCGGAGCTAA
site_3_destroyed
site_5_destroyed

entrez_gene

aliases	Bcl-2, PPP1R50
gene	BCL2
id	596

genbank_ids

mutation

name

Bcl-2

shRNA_sequence

size

species

9606

Homo sapiens