Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free (AB177817)

This data was developed using ab32396, the same antibody clone in a different buffer formulation.

Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling Estrogen Receptor alpha (phospho S118) with unpurified ab32396 at 5 μg/ml (1/200 dilution). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, an AlexaFluor^®488 Goat anti-Rabbit  was used as the secondary antibody at 2 μg/ml (1/1000 dilution). ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain at 2.5 μg/ml (1/200 dilution). DAPI nuclear counterstain.
Confocal image showing the signal increased after EGF (100ng/ml, 5 min) treatment and decreased after Lambda Protein Phosphatase treatment (31°C for 2 hours).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free (AB177817)

This data was developed using ab32396, the same antibody clone in a different buffer formulation.

Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) treated with EGF (100ng/ml, 5min) and treated with Lambda Protein Phosphatase 31°C for 2h cells labeling Estrogen receptor alpha (phospho S118) with purified ab32396 at 1 : 200 dilution (8.9μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1 : 1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free (AB177817)

This data was developed using ab32396, the same antibody clone in a different buffer formulation.

Immunofluorescent staining of (A) untreated and (B) Phosphatase-treated MCF-7 cells using unpurified ab32396.

• WB

Supplier Data

Western blot - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free (AB177817)

This data was developed using ab32396, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (<a href='/en-us/products/primary-antibodies/estrogen-receptor-alpha-phospho-s118-antibody-e91-ab32396'>ab32396</a>) at 1/1000 dilution

Lane 1:

MCF7 cell lysate (untreated)

Lane 2:

MCF7 cell lysate (treated with b-Estradiol and EGF)

Predicted band size: 66 kDa

Observed band size: 60 kDa

false

• WB

Supplier Data

Western blot - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free (AB177817)

This data was developed using ab32396, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (<a href='/en-us/products/primary-antibodies/estrogen-receptor-alpha-phospho-s118-antibody-e91-ab32396'>ab32396</a>) at 1/1000 dilution

Lane 1:

MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

MCF-7 (Human breast adenocarcinoma epithelial cell) treated with epidermal growth factor whole cell lysates at 15 µg

Lane 3:

MCF-7 (Human breast adenocarcinoma epithelial cell) treated with epidermal growth factor whole cell lysates, then the membrane was incubated with phosphatase at 15 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: 66 kDa

Observed band size: 60 kDa

false

Exposure time: 5s

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free (AB177817)

This data was developed using the same antibody clone in a different buffer formulation (ab32396).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 MCF7 (Human breast adenocarcinoma epithelial cell) cells treated with phenol red free medium and 5% charcoal stripped FBS for 3 days than treated with β-estradiol (10 nM 45 min) and 5 µg of ab32396 [E91]. 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-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free (AB177817)

This data was developed using the same antibody clone in a different buffer formulation (ab32396).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 MCF7 (Human breast adenocarcinoma epithelial cell) cells treated with phenol red free medium and 5% charcoal stripped FBS for 3 days than treated with β-estradiol (10 nM 45 min) and 5 µg of ab32396 [E91]. 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-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free (AB177817)

This data was developed using the same antibody clone in a different buffer formulation (ab32396).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 MCF7 (Human breast adenocarcinoma epithelial cell) cells treated with phenol red free medium and 5% charcoal stripped FBS for 3 days than treated with β-estradiol (10 nM 45 min) and 5 µg of ab32396 [E91]. 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.

• Dot

Unknown

Dot Blot - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] - BSA and Azide free (AB177817)

This data was developed using ab32396, the same antibody clone in a different buffer formulation.

Dot blot analysis of Lane 1 : Estrogen Receptor alpha (pS118) phospho peptide and Lane 2 : Estrogen Receptor alpha non-phospho peptide labeling Estrogen Receptor alpha (phospho S118) with unpurified ab32396 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time : 3 minutes.