Anti-PTEN antibody [SP218] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228466).

Flow cytometry overlay histogram showing wild-type Hap1 (green line) and PTEN knockout Hap1 stained with ab228466 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab228466) (1x 106 in 100μl at 0.04 μg/ml (1/55500)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type Hap1 - black line, PTEN knockout Hap1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in Hap1 Fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Intracellular Flow Cytometry analysis of MCF7 (human breast adenocarcinoma epithelial cell) cells labeling PTEN with purified ab228466 at 1/240 dilution (1.01μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Intracellular flow cytometric analysis of A431 (human epidermoid carcinoma cell line) cells labeling PTEN with ab228466 at 1/200 dilution (green) compared with anegative control rabbit IgG (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Formalin-fixed, paraffin-embedded human prostate adenocarcinoma tissue stained for PTEN using ab228466 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).

• WB

Lab

Western blot - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228466).

Western blot : Anti-PTEN antibody [SP218] - C-terminal ab228466 staining at 1/400 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 47 kDa in Wild-type HeLa cell lysates with no signal observed at this size in PTEN knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PTEN antibody [SP218] - C-terminal (<a href='/ko/products/primary-antibodies/pten-antibody-sp218-c-terminal-ab228466'>ab228466</a>) at 1/400 dilution

Lane 1:

Wild-type HeLa cell lysates at 20 µg

Lane 2:

Western blot - Human PTEN knockout HeLa cell line (<a href='/ko/products/cell-lines/human-pten-knockout-hela-cell-line-ab255419'>ab255419</a>) at 20 µg

Lane 3:

MCF7 cell lysates at 20 µg

Lane 4:

LNCaP cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

• WB

Lab

Western blot - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228466).

Western blot : Anti-PTEN antibody [SP218] - C-terminal ab228466 staining at 1/400 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 50 kDa in Wild-type A549 cell lysates with no signal observed at this size in PTEN knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PTEN antibody [SP218] - C-terminal (<a href='/ko/products/primary-antibodies/pten-antibody-sp218-c-terminal-ab228466'>ab228466</a>) at 1/400 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

Western blot - Human PTEN knockout A549 cell line (<a href='/ko/products/cell-lines/human-pten-knockout-a549-cell-line-ab286704'>ab286704</a>) at 20 µg

Lane 3:

Wild-type HeLa ab255928 cell lysates at 20 µg

Lane 4:

Western blot - Human PTEN knockout HeLa cell line (<a href='/ko/products/cell-lines/human-pten-knockout-hela-cell-line-ab255419'>ab255419</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [SP218] - BSA and Azide free (AB243940)

Cancer cells from prostate adenocarcinoma (panel A) and colon adenocarcinoma (panel D) show total loss or diminished expression of PTEN when SP218 is used. In contrast, nuclear staining (panel B and C, arrows) in cancer cells is present for clones D4.3 and 138G6. Diffuse cytoplasmic and stromal staining are prominent for clones D4.3 and 138G6. But interestingly, the stromal cell components (fibroblasts, lymphocytes, and endothelial cells) are stained with similar intensities across all images. This suggests that there is non-specific staining in stromal and cancer area with clones D4.3 and 138G6 when compared with SP218.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab228466).