According to the data sheet, for Immunohistochemistry, it references (PMID: 21249129). I see a 1:25 dilution in the journal article referenced under immunocytochemistry. Is this the same dilution recommended for IHC? If not do you have a recommended dilution? Antigen retrieval: Do you have any information on AR? Citrate buffer or higher pH (EDTA)? Temperature? Time? Incubation: Is 1 hour at room temperature or overnight at 4C recommended?

This antibody was validated in IHC in the reference PMID 11050159. Generally we recommend using antigen retrieval with citrate buffer and incubating for 1 hour at RT. Some antigens may be tricky and require overnight incubation. I imagine the specific protocol can be found in the reference, however.

Can I get the recipe for RIPA buffer and the lysate preparation protocol for membrane bound proteins using NB300-164?

Here is both the recipe we use for RIPA and a good protocol for tissue lysate prep. RIPA buffer: 150 mM sodium chloride 1.0% NP-40 or Triton X-100 0.5% sodium deoxycholate 0.1% SDS (sodium dodecyl sulphate) 50 mM Tris, pH 8.0 RIPA buffer is also useful for whole cell extracts and membrane-bound proteins, and may be preferable to NP-40 or Triton X100-only buffers for extracting nuclear proteins. It will disrupt protein-protein interactions and may therefore be problematic for immunoprecipitations/pull down assays. Preparation of lysate from tissues Dissect the tissue of interest with clean tools, on ice preferably, and as quickly as possible to prevent degradation by proteases. Place the tissue in round-bottom microfuge tubes or Eppendorf tubes and immerse in liquid nitrogen to snap freeze. Store samples at -80C for later use or keep on ice for immediate homogenization. For a approximately 5 mg piece of tissue, add approximately 300 ul lysis buffer rapidly to the tube, homogenize with an electric homogenizer, rinse the blade twice with another 2x300 ul lysis buffer, then maintain constant agitation for 2 hours at 4C (e.g place on an orbital shaker in the fridge). Volumes of lysis buffer must be determined in relation to the amount of tissue present (protein extract should not be too diluted to avoid loss of protein and large volumes of samples to be loaded onto gels. The minimum concentration is 0.1 mg/ml, optimal concentration is 1-5 mg/ml). Centrifuge for 20 min at 12000 rpm at 4C in a microcentrifuge. Gently remove the tubes from the centrifuge and place on ice, aspirate the supernatant and place in a fresh tube kept on ice; discard the pellet.

I want to know the concentration of Bestrophin 1 antibody [NB300-164].

For our product NB300-164, this antibody is provided as unpurified ascites. Therefore, the antibody concentration has not been determined.

We are interested in your item, NB300-164 and would you please let us know the antibody concentration of it?

The total protein concentration of the ascites (Lot# B-3) is 11.7 mg/ml.

Bestrophin 1 Antibody (E6-6) - BSA Free (NB300-164): Novus Biologicals

Name: Bestrophin 1 Antibody (E6-6) - BSA Free
Brand: Novus Biologicals
SKU: NB300-164
Rating: 5 (1 reviews)

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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

Reactivity	Hu, Mu, Po, Ca, Pm, Rt(-)Species Glossary
Applications	WB, Func, ICC/IF, IHC, IP, Dual ISH-IHC, KO
Clone	E6-6
Clonality	Monoclonal
Host	Mouse
Conjugate	Unconjugated
Format	BSA Free
Concentration	1.0 mg/ml

Catalog# & Conjugate	Size	Price
Alexa Fluor 350 NB300-164AF350

Alexa Fluor 405 NB300-164AF405

Alexa Fluor 488 NB300-164AF488

Alexa Fluor 532 NB300-164AF532

Alexa Fluor 594 NB300-164AF594

Alexa Fluor 647 NB300-164AF647

Alexa Fluor 700 NB300-164AF700

Alexa Fluor 750 NB300-164AF750

Allophycocyanin NB300-164APC

Biotin NB300-164B

CoraFluor 1 NB300-164CL1

DyLight 350 NB300-164UV

DyLight 405 NB300-164V

DyLight 488 NB300-164G

DyLight 550 NB300-164R

DyLight 594 NB300-164DL594

DyLight 650 NB300-164C

DyLight 680 NB300-164FR

DyLight 755 NB300-164IR

FITC NB300-164F

HRP NB300-164H

Janelia Fluor 525 NB300-164JF525

Janelia Fluor 549 NB300-164JF549

Janelia Fluor 585 NB300-164JF585

Janelia Fluor 635 NB300-164JF635

Janelia Fluor 646 NB300-164JF646

Janelia Fluor 669 NB300-164JF669

PE NB300-164PE

PerCP NB300-164PCP

mFluor Violet 450 SE NB300-164MFV450

mFluor Violet 500 SE NB300-164MFV500

mFluor Violet 610 SE NB300-164MFV610

Catalog# & Formulation	Size	Price
BSA Free NB300-164SS

Azide and BSA Free NBP2-80593

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB300-164AF350
Alexa Fluor 405	NB300-164AF405
Alexa Fluor 488	NB300-164AF488
Alexa Fluor 532	NB300-164AF532
Alexa Fluor 594	NB300-164AF594
Alexa Fluor 647	NB300-164AF647
Alexa Fluor 700	NB300-164AF700
Alexa Fluor 750	NB300-164AF750
Allophycocyanin	NB300-164APC
Biotin	NB300-164B
CoraFluor 1	NB300-164CL1
DyLight 350	NB300-164UV
DyLight 405	NB300-164V
DyLight 488	NB300-164G
DyLight 550	NB300-164R
DyLight 594	NB300-164DL594
DyLight 650	NB300-164C
DyLight 680	NB300-164FR
DyLight 755	NB300-164IR
FITC	NB300-164F
HRP	NB300-164H
Janelia Fluor 525	NB300-164JF525
Janelia Fluor 549	NB300-164JF549
Janelia Fluor 585	NB300-164JF585
Janelia Fluor 635	NB300-164JF635
Janelia Fluor 646	NB300-164JF646
Janelia Fluor 669	NB300-164JF669
PE	NB300-164PE
PerCP	NB300-164PCP
mFluor Violet 450 SE	NB300-164MFV450
mFluor Violet 500 SE	NB300-164MFV500
mFluor Violet 610 SE	NB300-164MFV610

Bestrophin 1 Antibody (E6-6) - BSA Free

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Bestrophin 1 Antibody (E6-6) - BSA Free Summary

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Publications for Bestrophin 1 Antibody (NB300-164)(49)

Review for Bestrophin 1 Antibody (NB300-164) (1) 51

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Immunogen	Synthetic peptide conjugated to KLH corresponding to the C-terminus of human Bestrophin 1 (KDHMDPYWALENRDEAHS) [Uniprot: O76090]
Isotype	IgG1 Kappa
Clonality	Monoclonal
Host	Mouse
Gene	BEST1
Purity	Protein A or G purified
Innovator's Reward	Test in a species/application not listed above to receive a full credit towards a future purchase. Learn about the Innovator's Reward

Storage	Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Buffer	PBS
Preservative	0.02% Sodium Azide
Concentration	1.0 mg/ml
Purity	Protein A or G purified

Publications using NB300-164	Applications	Species
Li J, Qiu C, Wei Y et al. Human Amniotic Epithelial Stem Cell-Derived Retinal Pigment Epithelium Cells Repair Retinal Degeneration Frontiers in Cell and Developmental Biology 2021-09-28 [PMID: 34650985]
Sripathi SR, Hu MW, Turaga RC Et al. Epithelial to mesenchymal transition (EMT) of human stem cell-derived retinal pigment epithelium shares commonalities with malignancy-associated EMT: a proteomic analysis Molecular & cellular proteomics : MCP 2021-08-26 [PMID: 34455105] (WB, Human)	WB	Human
Afanasyeva TAV, Corral-Serrano JC, Garanto A et al. A look into retinal organoids: methods, analytical techniques, and applications Cellular and Molecular Life Sciences 2021-10-01 [PMID: 34420069]
Schustak J, Twarog M, Wu X et al. Mechanism of Nucleic Acid Sensing in Retinal Pigment Epithelium (RPE): RIG-I Mediates Type I Interferon Response in Human RPE Journal of Immunology Research 2021-06-18 [PMID: 34239945] (ICC/IF, Human)	ICC/IF	Human
Choudhury R, Bayatti N, Scharff R Et Al. FHL-1 interacts with human RPE cells through the alpha 5 beta 1 integrin and confers protection against oxidative stress Scientific reports 2021-07-08 [PMID: 34239032] (ICC/IF)	ICC/IF
Zhao Q, Kong Y, Kittredge A et al. Distinct expression requirements and rescue strategies for BEST1 loss- and gain-of-function mutations eLife 2021-06-01 [PMID: 34061021] (WB)	WB
Tsai YT, Li Y, Ryu J et al. Impaired cholesterol efflux in retinal pigment epithelium of individuals with juvenile macular degeneration American journal of human genetics 2021-04-21 [PMID: 33909993]
D'Antonio-Chronowska, A, D'Antonio, M Et al. In vitro Differentiation of Human iPSC-derived Retinal Pigment Epithelium Cells (iPSC-RPE). Bio Protoc 2019-12-20 [PMID: 33654959] (WB, Mouse)	WB	Mouse
Biswas P, Borooah S, Matsui H et al. Detection and validation of novel mutations in MERTK in a simplex case of retinal degeneration using WGS and hiPSC-RPEs model Human mutation 2020-11-30 [PMID: 33252167]
Bonilha, V L, Bell, B A Et al. Cellular Changes in Retinas From Patients With BEST1 Mutations. Front Cell Dev Biol 2020-11-07 [PMID: 33154968] (ICC/IF)	ICC/IF
SchAfer N, Wolf HN, Enzbrenner A et al. Properdin Modulates Complement Component Production in Stressed Human Primary Retinal Pigment Epithelium Cells Antioxidants (Basel) 2020-08-26 [PMID: 32859013] (IF/IHC, Human)	IF/IHC	Human
Mitrousis N, Hacibekiroglu S, Ho M et al. Hydrogel-mediated co-transplantation of retinal pigmented epithelium and photoreceptors restores vision in an animal model of advanced retinal degeneration Biomaterials 2020-07-01 [PMID: 32791386] (ICC/IF, Mouse)	ICC/IF	Mouse
Zhao Zhang, Bin Yan, Fei Gao, Qing Li, Xiaohong Meng, Peikai Chen, Lei Zhou, Wen Deng, Cheng Li, Weiyi Xu, Shuo Han, Hong Feng, Yaping Li, Junhui Chen, Zhengqin Yin, Can Liao, Hung-Fat Tse, Aimin Xu, Qizhou Lian PSCs Reveal PUFA-Provoked Mitochondrial Stress as a Central Node Potentiating RPE Degeneration in Bietti’s Crystalline Dystrophy Molecular Therapy 2020-12-02 [PMID: 32755565]
Florian Udry, Sarah Decembrini, David M. Gamm, Nicole Déglon, Corinne Kostic, Yvan Arsenijevic Lentiviral mediated RPE65 gene transfer in healthy hiPSCs-derived retinal pigment epithelial cells markedly increased RPE65 mRNA , but modestly protein level Scientific Reports 2020-06-01 [PMID: 32483256]
Mladenov N, Petrova SD, Mladenova K et al. Miscibility of hBest1 and sphingomyelin in surface films - A prerequisite for interaction with membrane domains Colloids Surf B Biointerfaces 2020-02-21 [PMID: 32113084] (WB)	WB
Voisin A, Monville C, Plancheron A, et al. Cathepsin B pH-Dependent Activity Is Involved in Lysosomal Dysregulation in Atrophic Age-Related Macular Degeneration Oxid Med Cell Longev 2019-12-06 [PMID: 31885803] (Human)		Human
Ji C, Li Y, Kittredge A, et al. Investigation and Restoration of BEST1 Activity in Patient-derived RPEs with Dominant Mutations Sci Rep 2019-12-13 [PMID: 31836750] (WB, IF/ICC, Human)	WB, IF/ICC	Human
Smith E, D'Antonio-Chronowska A, Greenwald W et al. Human iPSC-derived retinal pigment epithelium: a model system for identifying and functionally characterizing causal variants at AMD risk loci Stem Cell Reports 2019-05-14 [PMID: 31080113]
Shahi PK, Hermans D, Sinha D et al. Gene Augmentation and Readthrough Rescue Channelopathy in an iPSC-RPE Model of Congenital Blindness Am. J. Hum. Genet. 2019-02-07 [PMID: 30686507] (ICC/IF, Human)	ICC/IF	Human
Zhang Y, Kittredge A, Ward N et al. ATP activates bestrophin ion channels through direct interaction Nat Commun 2018-08-07 [PMID: 30087350] (ICC/IF, Human)	ICC/IF	Human
Voisin A, Monville C, Plancheron A et al. HRPE cells derived from induced pluripotent stem cells are more sensitive to ozidative strss than ARPE-19 cell. Exp. Eye Res. 2018-07-23 [PMID: 30048622] (IF/IHC, Human)	IF/IHC	Human
Ramachandra Rao S, Pfeffer BA, MAs GOmez N et al. Compromised phagosome maturation underlies RPE pathology in cell culture and whole animal models of Smith-Lemli-Opitz Syndrome Autophagy. 2018-09-16 [PMID: 29979914] (WB, Human)	WB	Human
Li Y, Zhang Y, Xu Y et al. Patient-specific mutations impair BESTROPHIN1's essential role in mediating Ca(2+)-dependent Cl(-) currents in human RPE. Elife. 2017-10-24 [PMID: 29063836] (Human)		Human
Zhang Y, Zhang D, Wei W et al. Effects of RPE-conditioned medium on the differentiation of hADSCs into RPE cells, and their proliferation and migration. Exp Ther Med. 2017-10-01 [PMID: 29042966] (ICC/IF, Human)	ICC/IF	Human
Klingeborn M, Dismuke WM, Skiba NP et al. Directional Exosome Proteomes Reflect Polarity-Specific Functions in Retinal Pigmented Epithelium Monolayers Sci Rep 2017-07-07 [PMID: 28687758] (ICC/IF, Porcine)	ICC/IF	Porcine
Shim SH, Kim G, Lee DR et al. Survival of Transplanted Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells in a Human Recipient for 22 Months. JAMA Ophthalmol. 2017-02-09 [PMID: 28196199]
Kaneko Y, Pappas C, Tajiri N, Borlongan CV. Oxytocin modulates GABAAR subunits to confer neuroprotection in stroke in vitro. Sci Rep 2016-10-21 [PMID: 27767042]
Uggenti C, Briant K, Streit AK et al. Restoration of mutant bestrophin-1 expression, localisation and function. Dis Model Mech. 2016-08-12 [PMID: 27519691] (WB, ICC/IF, Human, Canine)	WB, ICC/IF	Human, Canine
Parker J, Mitrousis N, Shoichet MS. Hydrogel for Simultaneous Tunable Growth Factor Delivery and Enhanced Viability of Encapsulated Cells in Vitro. Biomacromolecules. 2016-02-08 [PMID: 26762290] (ICC/IF, Human)	ICC/IF	Human
Bertolotti E, Neri A, Camparini M et al. Stem cells as source for retinal pigment epithelium transplantation. Prog Retin Eye Res. 2014-06-13 [PMID: 24933042] (ICC/IF, Human) Details: Figure 4. Basolateral localization of bestrophin 1 (antibody used 1:1000, Novus Biologicals) in human RPE cells. Human retinal stem cells (RNS; derived from adult ciliary bodies of post-mortem donors) were differentiated into RPE and the molecular characteristics of RPE cells were evaluated - polarization of the RPE cells was assessed with three markers: bestrophin 1 at the basolateral membrane, and beta5-integrin as well as ezrin at the apical side of the RPE.	ICC/IF	Human
Sharon D, Al-Hamdani S, Engelsberg K et al. Ocular phenotype analysis of a family with biallelic mutations in the BEST1 gene. Am J Ophthalmol 2013-12-14 [PMID: 24345323] (ICC/IF, IHC-P, Human)	ICC/IF, IHC-P	Human
Doumanov J A, Zeitz C et al. Disease-causing mutations in BEST1 gene are associated with altered sorting of bestrophin-1 protein. Int J Mol Sci 2013-07-22 [PMID: 23880862] (ICC/IF, WB, Rat, Canine)	ICC/IF, WB	Rat, Canine
Maruotti J, Wahlin K, Gorrell D et al. A simple and scalable process for the differentiation of retinal pigment epithelium from human pluripotent stem cells. Stem Cells Transl Med 2013-05-01 [PMID: 23585288]
Blenkinsop TA, Salero E, Stern JH, Temple S. The culture and maintenance of functional retinal pigment epithelial monolayers from adult human eye Methods Mol Biol 2013-01-01 [PMID: 23097100]
Cho MS, Kim SJ, Ku SY et al. Generation of retinal pigment epithelial cells from human embryonic stem cell-derived spherical neural masses Stem Cell Res 2012-05-16 [PMID: 22683799] (ICC/IF, Human)	ICC/IF	Human
Ukrohne TU, Westenskow PD, Kurihara T et al. Generation of retinal pigment epithelial cells from small molecules and OCT4 reprogrammed human induced pluripotent stem cells. Stem Cells Transl Med 2012-02-01 [PMID: 22532929] (ICC/IF, Human)	ICC/IF	Human
Guziewicz KE, Slavik J, Lindauer SJ et al. Molecular consequences of BEST1 gene mutations in canine multifocal retinopathy predict functional implications for human bestrophinopathies. Invest Ophthalmol Vis Sci. 2011-04-15 [PMID: 21498618] (WB, Canine)	WB	Canine
Davidson AE, Millar ID, Burgess-Mullan R et al. Functional characterization of bestrophin-1 missense mutations associated with autosomal recessive bestrophinopathy. Invest Ophthalmol Vis Sci 2011-05-01 [PMID: 21330666]
Harness JV, Turovets NA, Seiler MJ et al. Equivalence of conventionally-derived and parthenote-derived human embryonic stem cells. PLoS ONE;6(1). 2011-01-01 [PMID: 21249129] (ICC/IF, WB, Human)	ICC/IF, WB	Human
Zhang Y, Patil RV, Marmorstein AD. Bestrophin 2 is expressed in human non-pigmented ciliary epithelium but not retinal pigment epithelium. Mol Vis;16:200-6. 2010-02-10 [PMID: 20157619] (WB, Human)	WB	Human
Feng Q, Lu SJ, Klimanskaya I et al. Hemangioblastic derivatives from human induced pluripotent stem cells exhibit limited expansion and early senescence. Stem Cells 2010-04-01 [PMID: 20155819] (ICC/IF, Human)	ICC/IF	Human
Davidson AE, Millar ID, Urquhart JE et al. Missense mutations in a retinal pigment epithelium protein, bestrophin-1, cause retinitis pigmentosa. Am J Hum Genet 2009-11-01 [PMID: 19853238] (Human)		Human
Buchholz DE, Hikita ST, RowlTJ et al. Derivation of functional retinal pigmented epithelium from induced pluripotent stem cells. Stem Cells 2009-10-01 [PMID: 19658190] (WB, Human)	WB	Human
Lu B, Malcuit C, Wang S et al. Long-term safety and function of RPE from human embryonic stem cells in preclinical models of macular degeneration. Stem Cells 2009-09-01 [PMID: 19521979] (WB, Human)	WB	Human
Guziewicz KE, Zangerl B, Lindauer SJ, Mullins RF, Sandmeyer LS, Grahn BH, Stone EM, AclGM, Aguirre GD. Bestrophin gene mutations cause canine multifocal retinopathy: a novel animal model for best disease. Invest Ophthalmol Vis Sci;48(5):1959-67. 2007-05-01 [PMID: 17460247] (WB, ICC/IF, Canine)	WB, ICC/IF	Canine
Mullins RF, Oh KT, Heffron E et al. Late development of vitelliform lesions and flecks in a patient with best disease: clinicopathologic correlation. Arch Ophthalmol 2005-11-01 [PMID: 16286623]
Klimanskaya, I, Hipp, J, Rezai, K et al. Derivation and comparative assessment of retinal pigment epithelium from human embryonic stem cells using transcriptomics. Cloning Stem Cells;6(3):217-45. 2004-01-01 [PMID: 15671670] (ICC/IF, Human)	ICC/IF	Human
Marmorstein LY, McLaughlin PJ, Stanton JB, Yan L, Crabb JW, Marmorstein AD. Bestrophin interacts physically and functionally with protein phosphatase 2A. J Biol Chem;277(34):30591-7. 2002-08-23 [PMID: 12058047] (WB, Porcine)	WB	Porcine
Marmorstein AD, Marmorstein LY, Rayborn M, Wang X, Hollyfield JG, Petrukhin K. Bestrophin, the product of the Best vitelliform macular dystrophy gene (VMD2), localizes to the basolateral plasma membrane of the retinal pigment epithelium. Proc Natl Acad Sci U S A;97(23):12758-63. 2000-11-07 [PMID: 11050159] (IP, WB, IHC-Fr, ICC/IF, Primate, Porcine, Human)	IP, WB, IHC-Fr, ICC/IF	Primate, Porcine, Human
Show All 49 Publications. Collapse Publications.

	Verified Customer 12/29/2016
Application:	WB
Species:	Human and Rat

Gene Symbol	BEST1
Entrez	7439(Human) 293735(Rat)
Uniprot	O76090(Human)