Anti-PTEN antibody [EPR22636-122] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stroma of human endometrial cancer (PMID : 2230170). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stroma of human ovarian cancer (PMID : 25608477). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling PTEN with ab267787 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human pancreas (PMID : 11021813). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

• IP

Unknown

Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

PTEN was immunoprecipitated from 0.35 mg MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate with ab267787 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267787 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2 : ab267787 IP in MCF7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab267787 in MCF7 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 15 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

All lanes:

Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] (<a href='/en-us/products/primary-antibodies/pten-antibody-epr22636-122-ab267787'>ab267787</a>)

Predicted band size: 47 kDa

Observed band size: 54 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

PTEN was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with ab267787 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267787 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2 : ab267787 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab267787 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 15 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

All lanes:

Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] (<a href='/en-us/products/primary-antibodies/pten-antibody-epr22636-122-ab267787'>ab267787</a>)

Predicted band size: 47 kDa

Observed band size: 54 kDa

false

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling PTEN with ab267787 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse pancreas (PMID : 11021813). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat pancreas (PMID : 11021813). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

• WB

Unknown

Western blot - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times :

Lanes 1-4 : 3 seconds;
Lane 5 : 114 seconds.

ab267787 was shown to specifically react with PTEN in wild-type HAP1 cells as signal was lost in PTEN knockout cells. Wild-type and PTEN knockout samples were subjected to SDS-PAGE. ab267787 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/20,000 dilution for 1 hour at room temperature before imaging.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 10514407).

Negative control : MDA-MB-468 (PMID : 21358673,15674339).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

All lanes:

Western blot - Anti-PTEN antibody [EPR22636-122] (<a href='/en-us/products/primary-antibodies/pten-antibody-epr22636-122-ab267787'>ab267787</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (Human near-haploid cell line) whole cell lysate at 20 µg

Lane 2:

PTEN knockout HAP1 whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

MDA-MB-468 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 54 kDa

false

• WB

Unknown

Western blot - Anti-PTEN antibody [EPR22636-122] - BSA and Azide free (AB267791)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab267787).

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times :

Lanes 1-6 : 3 seconds;
Lane 7 : 7 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID : 10514407).

All lanes:

Western blot - Anti-PTEN antibody [EPR22636-122] (<a href='/en-us/products/primary-antibodies/pten-antibody-epr22636-122-ab267787'>ab267787</a>) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

Lane 5:

Mouse kidney tissue lysate at 10 µg

Lane 6:

Mouse spleen tissue lysate at 10 µg

Lane 7:

Rat lung tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 54 kDa

false