Anti-PDX1 antibody [RM1269]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [RM1269] (AB323218)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling PDX1 with ab323218 at 1/100 (5.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human kidney. The section was incubated with ab323218 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PDX1 antibody [RM1269] (AB323218)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2(human hepatocellular carcinoma epithelial cell, Left) / DLD-1(human colorectal adenocarcinoma epithelial cell, Right) cells labelling PDX1 with ab323218 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : HepG2.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [RM1269] (AB323218)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling PDX1 with ab323218 at 1/100 (5.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human pancreas. The section was incubated with ab323218 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PDX1 antibody [RM1269] (AB323218)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized DLD-1(human colorectal adenocarcinoma epithelial cell) cells labelling PDX1 with ab323218 at 1/50 (10.5 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing mainly nuclear staining in DLD-1 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : HepG2.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [RM1269] (AB323218)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling PDX1 with ab323218 at 1/2000 (0.263 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse pancreas. The section was incubated with ab323218 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PDX1 antibody [RM1269] (AB323218)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3(mouse embryonic fibroblast, Left) / Beta-TC-6(mouse pancreas insulinoma beta cell, Right) cells labelling PDX1 with ab323218 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : NIH/3T3.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PDX1 antibody [RM1269] (AB323218)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Beta-TC-6 (mouse pancreas insulinoma beta cell) cells labelling PDX1 with ab323218 at 1/1000 (0.525 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing nuclear staining in Beta-TC-6 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : NIH/3T3 (PMID : 25271154).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [RM1269] (AB323218)

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling PDX1 with ab323218 at 1/2000 (0.263 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat pancreas. The section was incubated with ab323218 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

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Immunoprecipitation - Anti-PDX1 antibody [RM1269] (AB323218)

PDX1 was immunoprecipitated from 0.35 mg Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate with ab323218 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323218 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate
Lane 2 : ab323218 IP in Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab323218 in Beta-TC-6 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-PDX1 antibody [RM1269] (ab323218) at 1/30 dilution

All lanes:

Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 8s

• WB

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Western blot - Anti-PDX1 antibody [RM1269] (AB323218)

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

Negative control : HepG2, WI-38

The molecular weight observed is consistent with what has been described in the literature (PMID : 12488243, PMID : 12947327, PMID : 16732298)

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PDX1 antibody [RM1269] (ab323218) at 1/1000 dilution

Lane 1:

DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

WI-38 (human fetal lung fibroblast) whole cell lysate at 40 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

Observed band size: 46 kDa,36 kDa

true

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-PDX1 antibody [RM1269] (AB323218)

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

Negative control : NIH/3T3 (PMID : 25271154)

The molecular weight observed is consistent with what has been described in the literature (PMID : 12488243, PMID : 12947327, PMID : 16732298)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PDX1 antibody [RM1269] (ab323218) at 1/1000 dilution

Lane 1:

Beta-TC-6 (mouse pancreas insinoma beta cell) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) 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

Observed band size: 46 kDa,36 kDa

false

Exposure time: 37s