Anti-CCL25 antibody [EPR18002-133]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CCL25 antibody [EPR18002-133] (AB324594)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (Human embryonic kidney epithelial cell) transfected with a mouse CCL25 expression vector containing a myc-His-tag® cells labelling CCL25 with ab324594 at 1/1000 (0.513 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 positive staining in 293T transfected with a mouse CCL25 expression vector containing a myc-His-tag® (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 (0.38ug/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-CCL25 antibody [EPR18002-133] (AB324594)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a mouse IL-1RA expression CCL25 vector containing a myc-His-tag. (B) HEK-293T cells transfected with empty vector containing a myc-His-tag. tissue labeling CCL25 with ab324594 at 1/100 (5.13 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a mouse IL-1RA expression CCL25 vector containing a myc-His-tag. No staining on (B) HEK-293T cells transfected with empty vector containing a myc-His-tag.
The section was incubated with ab324594 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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCL25 antibody [EPR18002-133] (AB324594)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling CCL25 with ab324594 at 1/100 (5.13 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse colon (PMID : 10602049).
The section was incubated with ab324594 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCL25 antibody [EPR18002-133] (AB324594)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling CCL25 with ab324594 at 1/100 (5.13 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse liver (PMID : 9285413).
The section was incubated with ab324594 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCL25 antibody [EPR18002-133] (AB324594)

Immunohistochemical analysis of paraffin-embedded Mouse thymus tissue labeling CCL25 with ab324594 at 1/100 (5.13 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse thymus (PMID : 10602049, PMID : 9285413 ).
The section was incubated with ab324594 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCL25 antibody [EPR18002-133] (AB324594)

Immunohistochemical analysis of paraffin-embedded Mouse small intestine  tissue labeling CCL25 with ab324594 at 1/100 (5.13 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse small intestine (PMID : 9285413, PMID : 9285413 ).
The section was incubated with ab324594 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

• WB

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Western blot - Anti-CCL25 antibody [EPR18002-133] (AB324594)

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

Low expression : liver, kidney, spleen, lung (PMID : 9285413; PMID : 10602049)

The identity of the bands higher than 37 kDa in lane 3 and lane 6 are unknown.

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

All lanes:

Western blot - Anti-CCL25 antibody [EPR18002-133] (ab324594) at 1/1000 dilution

Lane 1:

Mouse thymus tissue lysate at 20 µg

Lane 2:

Mouse small intestine tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Lane 5:

Mouse spleen tissue lysate at 20 µg

Lane 6:

Mouse lung tissue 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: 14-16 kDa,36 kDa

false

Exposure time: 180s