Product datasheet
Anti-14-3-3 antibody ab9063
4 Abreviews 10 References 3 Images
Overview
Product name
Anti-14-3-3 antibody
Description
Rabbit polyclonal to 14-3-3
Specificity
Recognises all 14-3-3 isoforms.
Tested applications
IP, WB, ICC/IF
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: all Mammals
Immunogen
A mixture of 14-3-3 fusion proteins (beta, eta, tau, and sigma).
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.
Storage buffer
0.01M Na phosphate, 0.25M NaCl, pH7.6, 0.05% Na azide, 0.01% Thimerisol, 15mg/ml BSA
Purity
Whole antiserum
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab9063 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Notes
IP
Use at an assay dependent dilution.
WB
1/1000.
ICC/IF
Use a concentration of 5 µg/ml.
Target
Function
Adapter protein implicated in the regulation of a large spectrum of both general and specialized
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signaling pathways. Binds to a large number of partners, usually by recognition of a
phosphoserine or phosphothreonine motif. Binding generally results in the modulation of the
activity of the binding partner. Negative regulator of osteogenesis. Blocks the nuclear
translocation of the phosphorylated form (by AKT1) of SRPK2 and antagonizes its stimulatory
effect on cyclin D1 expression resulting in blockage of neuronal apoptosis elicited by SRPK2.
Sequence similarities
Belongs to the 14-3-3 family.
Post-translational
modifications
The alpha, brain-specific form differs from the beta form in being phosphorylated (By similarity).
Phosphorylated on Ser-60 by protein kinase C delta type catalytic subunit in a sphingosinedependent fashion.
Isoform Short contains a N-acetylmethionine at position 1.
Cellular localization
Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I
to stage IV.
Anti-14-3-3 antibody images
All lanes : Anti-14-3-3 antibody (ab9063) at
1/1000 dilution
Lane 1 : HeLa (Human epithelial carcinoma
cell line) Whole Cell Lysate
Lane 2 : Lung (Human) Tissue Lysate
Lane 3 : Brain (Human) Tissue Lysate - adult
normal tissue (ab29466)
Lysates/proteins at 10 µg per lane.
Western blot - 14-3-3 antibody (ab9063)
Secondary
Goat polyclonal to Rabbit IgG - H&L - PreAdsorbed (HRP) at 1/3000 dilution
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14-3-3 is required for CT-Raf kinase activity.
14-3-3 can be removed from CT-Raf in vitro
with the detergent Empigen-BB. Sf9 cells
were infected with recombinant baculoviruses
encoding either GST or a GST–CT-Raf fusion
protein. Forty-eight hours after infection, the
GST or GST–CT-Raf proteins were isolated
from NP-40 lysates by using glutathioneImmunoprecipitation - 14-3-3 antibody (ab9063)
Taken from Thorsen, J.A., et al, (1998) with
permission
agarose beads.
The bead-bound complexes were then
washed with either NP-40 lysis buffer (lanes 1
and 2) or NP-40 lysis buffer containing 1%
Empigen-BB (lane 3), resolved by SDSPAGE, transferred to nitrocellulose, and
developed with an antibody to either 14-3-3
(upper panel) or Raf (lower panel).
ICC/IF image of ab9063 stained PC12 cells.
The cells were 100% methanol fixed (5 min)
and then incubated in 1%BSA / 10% normal
goat serum / 0.3M glycine in 0.1% PBSTween for 1h to permeabilise the cells and
block non-specific protein-protein
interactions. The cells were then incubated
with the antibody (ab9063, 5µg/ml) overnight
at +4°C. The secondary antibody (green) was
Alexa Fluor® 488 goat anti-rabbit IgG (H+L)
used at a 1/1000 dilution for 1h. Alexa Fluor®
Immunocytochemistry/ Immunofluorescence-14-3-
594 WGA was used to label plasma
3 antibody(ab9063)
membranes (red) at a 1/200 dilution for 1h.
DAPI was used to stain the cell nuclei (blue)
at a concentration of 1.43µM.
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