Anti-Brachyury / Bry antibody [EPR18113]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

Immunohistochemical analysis of paraffin-embedded Human chordoma tissue labeling Brachyury / Bry with ab209665 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on Human chordoma is observed. Counter stained with Hematoxylin

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

Immunohistochemical analysis of paraffin-embedded Human chondrosarcoma tissue labeling Brachyury / Bry with ab209665 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Human chondrosarcoma. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

Immunohistochemical analysis of paraffin-embedded Human meningioma tissue labeling Brachyury / Bry with ab209665 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Human meningioma. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

Confocal image showing nucleus staining in MUG-Chor1 cell line (shown in green). The counterstain was observed in red. Nuclear DNA was labelled with DAPI (shown in blue).

ab209665 staining Brachyury/Bry in MUG-Chor1 (human sacral bone chordoma) cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Samples were incubated with primary antibody at a 1/1000 dilution.  An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at a 1/1000 dilution.  An Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used as a counterstain antibody.  DAPI was used as nuclear counterstain. Nuclear staining on MUG-Chor1 cells.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

ab209665 staining Brachyury/Bry in MUG-Chor1 (Human sacral bone chordoma) cell line by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and 90% methanol. Sample was incubated with primary antibody at 1/400 dilution (red). A Goat anti-rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution. Rabbit monoclonal IgG (ab172730) (Black) was used as an isotype control. Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Brachyury / Bry with ab209665 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Human kidney. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

Lane 1 (input) : MUG-Chor1 (human sacral bone chordoma) whole cell lysate, 10 μg
Lane 2 (+) : MUG-Chor1 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab209665 in MUG-Chor1 whole cell lysate

ab209665 immunoprecipitating Brachyury/Bry in MUG-Chor1 lysates. For western blotting, primary antibody used was ab209665 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking and diluting buffer used was 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Brachyury / Bry antibody [EPR18113] (ab209665)

Predicted band size: 47 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

Immunohistochemical analysis of formalin fixed paraffin embedded mouse embryo E14 labelling Brachyury/Bry with ab209665 at a concentration of 4µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a ChromoMap DAB IHC Kit with anti-rabbit HQ, anti-HQ HRP detection. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 64mins.

Anti-Brachyury/Bry antibody [EPR18113] ab209665 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-Brachyury / Bry antibody [EPR18113] (AB209665)

ab209665 staining Brachyury/Bry in Rat E14.5 embryo tissue sections by Immunohistochemistry (IHC-P). Tissue was fixed paraffin and antigen retrieval was performed by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody at a 1/4000 dilution. A ready to use Goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter-stain. Nuclear staining on notochord of rat E14.5 embryo.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

ab209665 staining Brachyury/Bry in mouse E14.5 embryo tissue sections by Immunohistochemistry (IHC-P). Tissue was fixed paraffin and antigen retrieval was performed by heat mediation using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody at a 1/4000 dilution. A ready to use Goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter-stain. Nuclear staining on notochord of mouse E14.5 embryo.

• WB

Lab

Western blot - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Brachyury / Bry antibody [EPR18113] (ab209665) at 1/1000 dilution

All lanes:

MUG-Chor1 (human sacral bone chordoma), whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 47 kDa

Observed band size: 43 kDa,49 kDa

false

Exposure time: 5s

• IHC - Wmt

Collaborator

IHC - Wholemount - Anti-Brachyury / Bry antibody [EPR18113] (AB209665)

IHC - Wholemount of E6.5 wholemount mouse embryo labelling Brachyury / Bry with ab209665. Sample was incubated with primary antibody (1/100 in PBS/0.1% Triton-X/5% donkey serum/1% BSA) for 18 hours at 4°C. An Alexa Fluor^® 488-conjugated donkey anti-rabbit IgG monoclonal was used as the secondary antibody (1/500).

This image is courtesy of an abreview by Dr Sophie Morgani