Anti-PDX1 antibody [EPR3358(2)]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized DLD-1(human colorectal adenocarcinoma epithelial cell) cells labelling PDX1 with ab134150 at 1/500 (2.05 µg/ml) dilution followed by a ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at a 1/1000 (2 µg) dilution (Green).

Confocal image showing nuclear staining in DLD-1 cells (shown in green), which is non-specific. 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 (2.5µg/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

ab134150 staining PDX1 in Human pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/500). An undiluted HRP-conjugated anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

ab134150, unpurified, staining PDX1 in Human pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% serum for 1 hour at 24°C; antigen retrieval was by heat mediation in 10mM sodium citrate, pH 6.0. Samples were incubated with primary antibody (1/1000 in 2% BSA) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

ab134150 staining PDX1 in the BXPC-3 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde. Samples were incubated with primary antibody (1/150). An Alexa Fluor^®555-conjugated Goat anti-rabbit IgG (1/500) was used as the secondary antibody. Nuclei were counterstained with DAPI.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

Immunohistochemical analysis of paraffin embedded normal Human Normal brain tissue using ab134150, unpurified, showing -ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

Immunohistochemical analysis of paraffin embedded Human duodenum tissue labelled with ab134150, unpurified, at 1/1000 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

Immunohistochemical analysis of paraffin embedded Human pancreas tissue labelled with ab134150, unpurified, at 1/1000 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

Immunohistochemical analysis of paraffin embedded Human Colonic adenocarcinoma tissue using ab134150, unpurified, showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

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Western blot - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

The expression profile observed in BxPC-3 is consistent with the literature (PMID : 12947327). Negative control : BxPC-3 (PMID : 12947327)

All lanes:

Western blot - Anti-PDX1 antibody [EPR3358(2)] (ab134150) at 1/1000 dilution

Lane 1:

Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Beta-TC-6 (Mouse pancreas insulinoma beta 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: 30 kDa

Observed band size: 40 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-PDX1 antibody [EPR3358(2)] (AB134150)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.