Anti-PMS2 antibody [EPR3947] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using ab110638, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma tissue sections labeling PMS2 with Purified ab214442 at 1 : 600 (2.71 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using ab110638, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling PMS2 with ab110638 at a concentration of 3µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

Anti-PMS2 antibody [EPR3947] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using ab214442, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling PMS2 with Purified ab214442 at 1 : 600 (2.71 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using ab110638, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue sections labeling PMS2 with Purified ab214442 at 1 : 600 (2.71 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using ab110638, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling PMS2 with Purified ab214442 at 1 : 200 dilution (8 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

Tissue Microarrays stained for "Anti-PMS2 antibody [EPR3947]" using "ab110638"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab110638 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using ab110638, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling PMS2 with Purified ab214442 at 1 : 160 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IP

Lab

Immunoprecipitation - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using ab110638, the same antibody clone in a different buffer formulation.
Purified ab110638 at 1/50 dilution (2μg) immunoprecipitating PMS2 in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab110638 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab110638 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 110 kDa
Lower bands are degradation bands and fresh lysate is recommended.

All lanes:

Immunoprecipitation - Anti-PMS2 antibody [EPR3947] (<a href='/en-us/products/primary-antibodies/pms2-antibody-epr3947-ab110638'>ab110638</a>)

Predicted band size: 96 kDa

false

• IP

Lab

Immunoprecipitation - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using ab110638, the same antibody clone in a different buffer formulation.
Purified ab110638 at 1/50 dilution (2μg) immunoprecipitating PMS2 in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab110638 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab110638 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 110 kDa
Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.

All lanes:

Immunoprecipitation - Anti-PMS2 antibody [EPR3947] (<a href='/en-us/products/primary-antibodies/pms2-antibody-epr3947-ab110638'>ab110638</a>)

Predicted band size: 96 kDa

false

• WB

Supplier Data

Western blot - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using ab110638, the same antibody clone in a different buffer formulation.

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

ab181602 GADPH was used as a loading control.

Lower bands are degradation bands and fresh lysate is recommended to get desired WB result.

All lanes:

Western blot - Anti-PMS2 antibody [EPR3947] (<a href='/en-us/products/primary-antibodies/pms2-antibody-epr3947-ab110638'>ab110638</a>) at 1/10000 dilution

Lane 1:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate, freshly made at 20 µg

Lane 2:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, freshly made at 20 µg

Lane 4:

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: 96 kDa

Observed band size: 120 kDa

false

Exposure time: 20s

• WB

Lab

Western blot - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This WB data was generated using the same anti-PMS2 antibody clone, EPR3947, in a different buffer formulation (cat# ab110638).

Lanes 1-2 : Merged signal (red and green). Green - ab110638 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab110638 was shown to specifically react with PMS2 in wild-type HAP1 cells. No band was observed when PMS2 knockout samples were used. Wild-type and PMS2 knockout samples were subjected to SDS-PAGE. ab110638 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PMS2 antibody [EPR3947] (<a href='/en-us/products/primary-antibodies/pms2-antibody-epr3947-ab110638'>ab110638</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 30 µg

Lane 2:

PMS2 knockout HAP1 whole cell lysate at 30 µg

Predicted band size: 96 kDa

false

• WB

Lab

Western blot - Anti-PMS2 antibody [EPR3947] - BSA and Azide free (AB214442)

This data was developed using the same antibody clone in a different buffer formulation (ab110638).

Lanes 1- 2 : Merged signal (red and green). Green - ab110638 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab110638 was shown to react with PMS2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261776 (knockout cell lysate ab257142) was used. Wild-type HeLa and PMS2 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. ab110638 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 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-PMS2 antibody [EPR3947] (<a href='/en-us/products/primary-antibodies/pms2-antibody-epr3947-ab110638'>ab110638</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PMS2 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-pms2-knockout-hela-cell-lysate-ab257142'>ab257142</a>) at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 96 kDa

Observed band size: 120 kDa,37 kDa

false