Anti-PTEN antibody [Y184]

• Supplier: Abcam
• Catalog: ab32199
• Prices: $401.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: Y184
• Applications: WB FC IP ICC/IF ICC/IF
• Species Reactivities: Mouse, Rat, Human
• Antigen: A synthetic peptide corresponding to residues in the C-term of human PTEN
• Blocking Peptide: PTEN peptide
• Description: Rabbit Monoclonal
• Gene: PTEN
• Conjugate: Unconjugated

Product images

Anti-PTEN antibody [Y184] (ab32199) at 1/500 dilution + MCF7 cell lysate

ab32199 staining PTEN in human white blood cells by Immunocytochemistry/ Immunofluorescence. The cells were acetone fixed then blocked using 1% BSA for 1 hour at 25°C. Samples were then incubated with primary antibody at a 1/100 dilution for 2 hours at 25°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/500 dilution. DAPI was used to stain the cell nuclei (blue).See Abreview

Overlay histogram showing MCF-7 cells stained with ab32199 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32199, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

ICC/IF image of ab32199 stained HepG2 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 (ab32199, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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.

All lanes : Anti-PTEN antibody [Y184] (ab32199) at 1/500 dilutionLane 1 : Mouse primary bone marrow derived macrophage whole cell lysateLane 2 : Mouse primary bone marrow derived macrophage whole cell lysateLane 3 : Mouse primary bone marrow derived macrophage whole cell lysateLane 4 : Mouse primary bone marrow derived macrophage whole cell lysateLysates/proteins at 50 µg per lane.SecondaryHRP-conjugated goat anti-rabbit IgG polyclonaldeveloped using the ECL techniquePerformed under reducing conditions.

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