Validation Data Gallery

Filter: AllWBIPIHCIF/ICCFC (Intra)

WB analysis using 14503-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 14503-1-AP (14-3-3 antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.
WB analysis of mouse heart using 14503-1-AP
mouse heart tissue were subjected to SDS PAGE followed by western blot with 14503-1-AP (14-3-3 Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.
WB analysis of Jurkat using 14503-1-AP
Jurkat cells were subjected to SDS PAGE followed by western blot with 14503-1-AP (14-3-3 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
WB analysis of rat liver using 14503-1-AP
rat liver tissue were subjected to SDS PAGE followed by western blot with 14503-1-AP (14-3-3 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
WB analysis of mouse liver using 14503-1-AP
mouse liver tissue were subjected to SDS PAGE followed by western blot with 14503-1-AP (14-3-3 Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.
WB analysis of mouse brain using 14503-1-AP
mouse brain tissue were subjected to SDS PAGE followed by western blot with 14503-1-AP (14-3-3 Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.
IP experiment of mouse lung using 14503-1-AP
IP result of anti-14-3-3 (IP:14503-1-AP, 5ug; Detection:14503-1-AP 1:500) with mouse lung tissue lysate 4000ug.
IHC staining of mouse brain using 14503-1-AP
Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 14503-1-AP (14-3-3 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human testis using 14503-1-AP
Immunohistochemical analysis of paraffin-embedded human testis tissue slide using 14503-1-AP (14-3-3 Antibody) at dilution of 1:50 (under 10x lens).
IHC staining of human testis using 14503-1-AP
Immunohistochemical analysis of paraffin-embedded human testis tissue slide using 14503-1-AP (14-3-3 Antibody) at dilution of 1:50 (under 40x lens).
IF Staining of HepG2 using 14503-1-AP
Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using 14-3-3 antibody (14503-1-AP) at dilution of 1:400 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).
IF Staining of HepG2 using 14503-1-AP
Immunofluorescent analysis of HepG2 cells, using YWHAQ antibody 14503-1-AP at 1:50 dilution and Rhodamine-labeled goat anti-rabbit IgG (red).
FC experiment of HeLa using 14503-1-AP
1x10^6 HeLa cells were intracellularly stained with 0.4 ug 14-3-3 Polyclonal antibody (14503-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.4 ug rabbit IgG isotype control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer.

Featured Product
KD/KO Validated

14-3-3 Polyclonal antibody

14-3-3 Polyclonal Antibody for WB, IF/ICC, FC (Intra), IP, ELISA

Cat No. 14503-1-AP

Publications(18)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), IP, CoIP, RIP, ELISA

Synonyms

YWHAQ, 14-3-3 protein T-cell, 14-3-3 theta, 14-3-3 zeta, 14-3-3σ

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

CoraLite® Plus 555 CoraLite® Plus 647

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (18)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	HeLa cells, mouse liver tissue, mouse brain tissue, Jurkat cells, rat liver tissue, mouse heart tissue, NIH/3T3 cells, C6 cells
Positive IP detected in	mouse lung tissue
Positive IF/ICC detected in	HepG2 cells
Positive FC (Intra) detected in	HeLa cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:1000-1:4000
Immunoprecipitation (IP)	IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunofluorescence (IF)/ICC	IF/ICC : 1:200-1:800
Flow Cytometry (FC) (INTRA)	FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Published Applications

KD/KO	See 3 publications below
WB	See 14 publications below
IHC	See 2 publications below
IF	See 1 publications below
IP	See 4 publications below
CoIP	See 3 publications below
RIP	See 2 publications below

Product Information

14503-1-AP targets 14-3-3 in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, RIP, ELISA applications and shows reactivity with human, mouse, rat samples.

Tested Reactivity	human, mouse, rat
Cited Reactivity	human, mouse, rat
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag5959 Product name: Recombinant human 14-3-3 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 1-245 aa of BC056867 Sequence: MEKTELIQKAKLAEQAERYDDMATCMKAVTEQGAELSNEERNLLSVAYKNVVGGRRSAWRVISSIEQKTDTSDKKLQLIKDYREKVESELRSICTTVLELLDKYLIANATNPESKVFYLKMKGDYFRYLAEVACGDDRKQTIDNSQGAYQEAFDISKKEMQPTHPIRLGLALNFSVFYYEILNNPELACTLAKTAFDEAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDSAGEECDAAEGAEN 相同性解析による交差性が予測される生物種
Full Name	tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, theta polypeptide
Calculated molecular weight	28 kDa
Observed molecular weight	31 kDa
GenBank accession number	BC056867
Gene Symbol	14-3-3 theta
Gene ID (NCBI)	10971
RRID	AB_2218096
Conjugate	Unconjugated
Form
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	P27348
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

14-3-3 proteins are the first phosphoserine/phosphothreonine-binding proteins to be discovered. 14-3-3 family members interact with a wide spectrum of proteins and possess diverse functions. Mammals express seven distinct 14-3-3 isoforms (gamma, epsilon, beta, zeta, sigma, theta, tau) that form multiple homo- and hetero- dimmers. 14-3-3 proteins display the highest expression levels in the brain, and have been implicated in several neurodegenerative diseases, including Alzheimer's disease and amyotrophic lateral sclerosis.

Protocols

Product Specific Protocols
IF protocol for 14-3-3 antibody 14503-1-AP	Download protocol
IHC protocol for 14-3-3 antibody 14503-1-AP	Download protocol
IP protocol for 14-3-3 antibody 14503-1-AP	Download protocol
WB protocol for 14-3-3 antibody 14503-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
human	WB	Neuron Targeting 14-3-3θ-mediated TDP-43 pathology in amyotrophic lateral sclerosis and frontotemporal dementia mice Authors - Yazi D Ke View Article
		Int J Biol Macromol Biofilm formation initiating rotifer-specific biopolymer and its predicted components Authors - Zsolt Datki View Article
human	WB	Elife Ser/Thr kinase Trc controls neurite outgrowth in Drosophila by modulating microtubule-microtubule sliding. Authors - Rosalind Norkett KD Validated View Article
human	CoIP	Int J Mol Sci APEX2-Mediated Proximity Labeling Resolves the DDIT4-Interacting Proteome. Authors - Marianna Naki View Article
mouse,human	WB,IP	Aging (Albany NY) The interaction protein of SORBS2 in myocardial tissue to find out the pathogenic mechanism of LVNC disease. Authors - Chunyan Li View Article
human	WB	J Inflamm Res PADI6 Regulates Trophoblast Cell Migration-Invasion Through the Hippo/YAP1 Pathway in Hydatidiform Moles. Authors - Bo Huang View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis using 14503-1-AP

WB analysis of mouse heart using 14503-1-AP

WB analysis of Jurkat using 14503-1-AP

WB analysis of rat liver using 14503-1-AP

WB analysis of mouse liver using 14503-1-AP

WB analysis of mouse brain using 14503-1-AP

IP experiment of mouse lung using 14503-1-AP

IHC staining of mouse brain using 14503-1-AP

IHC staining of human testis using 14503-1-AP

IHC staining of human testis using 14503-1-AP

IF Staining of HepG2 using 14503-1-AP

IF Staining of HepG2 using 14503-1-AP

FC experiment of HeLa using 14503-1-AP

14-3-3 Polyclonal antibody

14-3-3 Polyclonal Antibody for WB, IF/ICC, FC (Intra), IP, ELISA

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

Targeting 14-3-3θ-mediated TDP-43 pathology in amyotrophic lateral sclerosis and frontotemporal dementia mice

Biofilm formation initiating rotifer-specific biopolymer and its predicted components

Ser/Thr kinase Trc controls neurite outgrowth in Drosophila by modulating microtubule-microtubule sliding.

APEX2-Mediated Proximity Labeling Resolves the DDIT4-Interacting Proteome.

The interaction protein of SORBS2 in myocardial tissue to find out the pathogenic mechanism of LVNC disease.

PADI6 Regulates Trophoblast Cell Migration-Invasion Through the Hippo/YAP1 Pathway in Hydatidiform Moles.