Anti-CDX2 antibody [EPR2764Y]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Immunohistochemical analysis of formalin fixed paraffin embedded human small intestine labelling CDX2 with ab76541 at a concentration of 0.05 µg/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.

ab76541 anti-CDX2 [EPR2764Y] 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

AbReview19013****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Unpurified ab76541 staining CDX2 in human intestinal tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

Tissue was fixed with paraformaldehyde and blocked with 5% BSA for 1 hours at 25°C; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/100 in PBS containing 5% BSA) for 18 hours at 4°C. An HRP-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SW480 ( human colorectal adenocarcinoma epithelial cell) labelling CDX2 with ab76541 at 1/400 (1.82 µg/ml) dilution (Green).

Negative control : MCF7.

Confocal image showing nuclear staining on the SW480 cell line.

Nuclear DNA was labelled with DAPI (shown in blue).

Secondary used was ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody -Microtubule Marker (Alexa Fluor® 594) was used as a counterstain at 1/200 (2.5µg/ml) dilution.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling CDX2 with Purified ab76541 at 1 : 1000 dilution (0.8 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Unpurified ab76541, at 1/250 dilution, staining CDX2 in paraffin-embedded human gastric adenocarcinoma tissue, by immunohistochemistry. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Unpurified ab76541 showing negative staining in normal kidney tissue. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Unpurified ab76541 showing negative staining in skeletal muscle tissue. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Unpurified ab76541 showing positive staining in normal colon tissue. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Unpurified ab76541 showing positive staining in colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Unpurified ab76541 showing negative staining in normal brain tissue. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Unpurified ab76541 showing negative staining in normal brain tissue. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling CDX2 with Purified ab76541 at 1 : 1000 dilution (0.8 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) ab97051
was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue sections labeling CDX2 with Purified ab76541 at 1 : 1000 dilution (0.8 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) ab97051
was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Lab

Western blot - Anti-CDX2 antibody [EPR2764Y] (AB76541)

All lanes:

Western blot - Anti-CDX2 antibody [EPR2764Y] (ab76541) at 1/10000 dilution

Lane 1:

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

Lane 2:

Human colon cancer lysates at 20 µg

Lane 3:

Rat colon lysates at 20 µg

Lane 4:

Mouse colon lysates at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/10000 dilution

Predicted band size: 33 kDa

Observed band size: 40 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-CDX2 antibody [EPR2764Y] (AB76541)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ Caco-2 (human colorectal adenocarcinoma epithelial cell) cells and 5 µg of ab76541 [EPR2764Y]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-CDX2 antibody [EPR2764Y] (AB76541)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ Caco-2 (human colorectal adenocarcinoma epithelial cell) cells and 5 µg of ab76541 [EPR2764Y]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-CDX2 antibody [EPR2764Y] (AB76541)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ Caco-2 (human colorectal adenocarcinoma epithelial cell) cells and 5 µg of ab76541 [EPR2764Y]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• WB

Unknown

Western blot - Anti-CDX2 antibody [EPR2764Y] (AB76541)

Lane 1:

Western blot - Anti-CDX2 antibody [EPR2764Y] (ab76541) at 1/1000 dilution

Lanes 2 - 3:

Western blot - Anti-CDX2 antibody [EPR2764Y] (ab76541) at 1/5000 dilution

Lane 1:

HT-29 (Human colorectal adenocarcinoma cell line) lysate at 10 µg

Lane 2:

Colon cancer lysate at 10 µg

Lane 3:

Human small intestine lysate at 10 µg

Secondary

All lanes:

HRP labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 33 kDa

Observed band size: 40 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-CDX2 antibody [EPR2764Y] (AB76541)

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.