Anti-PTEN antibody [EPR4408-76]

• WB

Lab

Western blot - Anti-PTEN antibody [EPR4408-76] (AB133532)

Lanes 1- 2 : Merged signal (red and green). Green - ab133532 observed at 47 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

ab133532 was shown to react with PTEN in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255419 (knockout cell lysate ab263829) was used. Wild-type HeLa and PTEN knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133532 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTEN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTEN knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-pten-knockout-hela-cell-line-ab255419'>ab255419</a>)

Predicted band size: 47 kDa

Observed band size: 47 kDa

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• WB

Unknown

Western blot - Anti-PTEN antibody [EPR4408-76] (AB133532)

Lanes 1 and 2 : Green signal from target - ab133532 observed at 47 kDa
Lanes 3 and 4 : Red signal from loading control - ab8245 observed at 37 kDa
Lanes 5 and 6 : Merged (red and green) signal

ab133532 was shown to specifically react with PTEN in wild-type HAP1 cells. No band was observed when PTEN knockout samples were used. Wild-type and PTEN knockout samples were subjected to SDS-PAGE, ab133532 and ab8245 (loading control to GAPDH) were diluted to 1/10,000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

Lanes 1 - 2:

Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532) at 1/10000 dilution

Lanes 3 - 4:

Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532) at 1/2000 dilution

Lanes 5 - 6:

Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)

Lanes 1, 3 and 5:

PTEN knockout HAP1 cell lysate at 20 µg

Lanes 2, 4 and 6:

Wild-type HAP1 cell lysate at 20 µg

Predicted band size: 47 kDa

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• WB

Lab

Western blot - Anti-PTEN antibody [EPR4408-76] (AB133532)

Lanes 1 - 3 : Merged signal (red and green). Green - ab133532 observed at 47 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab133532 was shown to react with PTEN in wild-type HeLa cells in western blot. The bands observed in PTEN knockout cell line ab255419 (PTEN knockout cell lysate ab263829) below 47 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. HeLa wild-type and PTEN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab133532 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTEN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTEN knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-pten-knockout-hela-cell-line-ab255419'>ab255419</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

• WB

Unknown

Western blot - Anti-PTEN antibody [EPR4408-76] (AB133532)

All lanes:

Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532) at 1/10000 dilution

Lane 1:

C6 cell lysate at 10 µg

Lane 2:

RAW 264.7 cell lysate at 10 µg

Lane 3:

NIH 3T3 cell lysate at 10 µg

Lane 4:

MCF7 cell lysate at 10 µg

Lane 5:

293T cell lysate at 10 µg

Lane 6:

A431 cell lysate at 10 µg

Lane 7:

HeLa cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 47 kDa

Observed band size: 54 kDa

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• WB

CiteAb

Western blot - Anti-PTEN antibody [EPR4408-76] (AB133532)

PTEN western blot using anti-PTEN antibody [EPR4408-76] ab133532. Publication image and figure legend from Zhong, C., Wang, K., et al., 2016, J Cell Mol Med, PubMed 27061862.

ab133532 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab133532 please see the product overview.

Pten is a target gene of miR-19b involved in proliferation. (A) miR-19b negatively regulated Pten at the protein level in cardiac fibroblasts. (B) Pten siRNAs decreased Pten at least at mRNA level. (C) Pten siRNAs significantly abolished the inhibitory effect of miR-19b in proliferation of cardiac fibroblasts. Scale bar, 100 μm. *p < 0.05 versus control; #p < 0.05 versus miR-19b inhibitor. Pten, Phosphatase and tensin homologue.

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