Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

Formalin-fixed, paraffin-embedded human breast tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab16660)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

Human breast carcinoma stained with ab16660.

This image was generated using the hybridoma version of the product.

.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab16660)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

IHC image of ab16660 staining Estrogen Receptor alpha in normal human cervix formalin-fixed paraffin-embedded tissue sections* performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6 epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16660 1/250 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

This image was generated using the hybridoma version of the product.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank supported by the NIHR Cambridge Biomedical Research Centre

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab16660)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

Formalin-fixed, paraffin-embedded human breast ductal carcinoma tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab16660)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

Formalin-fixed paraffin-embedded human breast carcinoma tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

This image was generated using the hybridoma version of the product.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab16660)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling ER alpha with ab187260 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 min with ULTRA cell conditioning solution (CC1 pH 8.5). ab187260 anti ER alpha antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

Formalin-fixed, paraffin-embedded human ovarian adenocarcinoma tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab16660)

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

This data was developed using ab16660, the same antibody clone in a different buffer formulation. Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human triple-positive breast carcinoma tissue sections labeling Estrogen Receptor (ER) with ab16660, at a 1/200 dilution ( 0.07 μg/ml). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins and Opal Polymer HRP Ms + Rb was used as the secondary antibody. DAPI was used as the nuclear counterstain. Panel A : merged staining of anti-Progesterone Receptor (PR) (magenta; Opal™690), anti-HER2 (red; Opal™570) and anti-Estrogen Receptor (ER) (green; Opal™520) on human triple-positive breast carcinoma. Panel B : anti-PR stained on nucleus of cancer cells. Panel C : anti-HER2 stained on membrane of cancer cells. Panel D : anti-ER stained on nucleus of cancer cells. The section was incubated in three rounds of staining : in the order of ab16661 for 30 mins, then ab16660 and ab231438 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

This data was developed using ab16660, the same antibody clone in a different buffer formulation. Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human mammary gland tissue sections labeling Estrogen Receptor (ER) with ab16660, at a 1/200 dilution ( 0.07 μg/ml). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins and Opal Polymer HRP Ms + Rb was used as the secondary antibody. DAPI was used as the nuclear counterstain. Panel A : merged staining of anti-Progesterone Receptor (PR) (magenta; Opal™690), anti-HER2 (red; Opal™570) and anti-Estrogen Receptor (ER) (green; Opal™520) on human mammary gland. Panel B : anti-PR stained on nucleus of some ductal cells. Panel C : anti-HER2 stained on no cells. Panel D : anti-ER stained on nucleus of some ductal cells. The section was incubated in three rounds of staining : in the order of ab16661 for 30 mins, then ab16660 and ab231438 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

This data was developed using ab16660, the same antibody clone in a different buffer formulation. Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human triple-negative breast carcinoma tissue sections labeling Estrogen Receptor (ER) with ab16660, at a 1/200 dilution ( 0.07 μg/ml). Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins and Opal Polymer HRP Ms + Rb was used as the secondary antibody. DAPI was used as the nuclear counterstain. Panel A : merged staining of anti-Progesterone Receptor (PR) (magenta; Opal™690), anti-HER2 (red; Opal™570) and anti-Estrogen Receptor (ER) (green; Opal™520) on human triple-negative breast carcinoma. Panel B : anti-PR stained on no cells. Panel C : anti-HER2 stained on no cells. Panel D : anti-ER stained on no cells. The section was incubated in three rounds of staining : in the order of ab16661 for 30 mins, then ab16660 and ab231438 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• WB

Lab

Western blot - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

This data was developed using ab16660, the same antibody clone in a different buffer formulation.

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 16165085, PMID : 31036290). The identity of the lower MW band below 37kDa is unknown.

All lanes:

Western blot - Anti-Estrogen Receptor alpha antibody [SP1] (<a href='/en-us/products/primary-antibodies/estrogen-receptor-alpha-antibody-sp1-ab16660'>ab16660</a>) at 1/1000 dilution

All lanes:

T-47D (human mammary gland ductal carcinoma epithelial cell) whole cell lysates 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: 46-66 kDa

false

Exposure time: 180s

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

ab16660 staining Estrogen Receptor alpha in MCF7 cells. The cells were fixed with 4% formaldehyde (10min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab16660 at 1/250 dilution (shown in green) and ab195889 Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 594) at 2μg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

This image was generated using the hybridoma version of the product.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab16660)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

This data was developed using ab16660, the same antibody clone in a different buffer formulation. Tissue Microarrays stained for Anti-Estrogen Receptor alpha antibody [SP1] using ab16660 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested. The section was incubated with ab16660 for 10 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

Intracellular flow cytometry analysis of MCF7 (human breast adenocarcinoma epithelial cell) labeling Estrogen Receptor alpha with purified ab16660 at 1/200 dilution (1.06 μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor^® 488 ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (black). Unlableled control - Unlabelled cells (blue).This image was generated using the hybridoma version of the product.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab16660).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [SP1] - BSA and Azide free (AB187260)

Clone SP1 (ab187260) has been successfully conjugated by Abcam. This image was generated using Anti-Estrogen Receptor alpha antibody [SP1] (Alexa Fluor® 647). Please refer to ab267512 for protocol details.

IHC image of Estrogen Receptor alpha staining in a section of formalin-fixed paraffin-embedded normal human breast*.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Biocare Medical NxGen pressure cooker using retrieval settings of 110°C for 20 minutes. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab267512 at 1/100 dilution (shown in red) and counterstained using ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

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

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.