Key features and details
- Rabbit polyclonal to YWHAE
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Cow, Human
- Isotype: IgG
Product nameAnti-YWHAE antibody
See all YWHAE primary antibodies
DescriptionRabbit polyclonal to YWHAE
SpecificityRaised against an N-terminal epitope which is N-acetylated as in the normal protein therefore may not react with recombinant protein expressed in E. coli that is not N-acetylated.
Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
Species reactivityReacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Cow, Human
Predicted to work with: Mammals
Synthetic peptide corresponding to YWHAE (N terminal).
This product was previously labelled as 14-3-3 epsilon
Storage instructionsShipped 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.
Concentration information loading...
Our Abpromise guarantee covers the use of ab43057 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration.|
|WB||1/3000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).|
FunctionAdapter protein implicated in the regulation of a large spectrum of both general and specialized signaling pathway. 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.
Sequence similaritiesBelongs to the 14-3-3 family.
Cellular localizationCytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
- Information by UniProt
- 14 3 3 E antibody
- 14 3 3 epsilon antibody
- 14 3 3E antibody
All lanes : Anti-YWHAE antibody (ab43057) at 1/500 dilution
Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 29 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?
Additional bands at: 19 kDa, 74 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 seconds
ICC/IF image of ab43057 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% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab43057, 1/1000 dilution) 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® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Ab43057 staining human normal placenta tissue. Staining is localised to cytoplasm.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system (DAKO Autostainer Plus ), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT Link. . Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
ab43057 has been referenced in 2 publications.