Anti-AP2 gamma/TFAP2C antibody [EPR20331]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Panel A : anti-SIGLEC6 (green; Opal™520), anti-TFAP2C (magenta; Opal™690) and anti-PLVAP/PV-1 (gray; Opal™570) on human placenta. Panel B : anti-SIGLEC6 staining membrane of trophoblast in human placenta. Panel C : anti-TFAP2C staining nucleus of trophoblast in human placenta. Panel D : anti-PLVAP/PV-1 staining endothelium in human placenta. Nuclear DNA was labeled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab317307, ab218107 and ab321889 for 30 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling AP2 gamma/TFAP2C with ab218107 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Nuclear staining on poorly differentiated human breast cancer is observed. The data is consistent with the literature (PMID : 18825690).

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human placenta tissue staining PAPP A2 with ab307087 at a 1/5000 dilution, ab218107 anti-TFAP2C used at 1/200 dilution and ab315802 anti-CD34 used at a 1/2000 dilution.

Panel A : anti-PAPP A2 (green; Opal™520), anti-TFAP2C (magenta; Opal™690) and anti-CD34 (gray; Opal™570) on human placenta.

Panel B : anti-PAPP A2 staining cytoplasm of trophoblast in human placenta.

Panel C : anti-TFAP2C staining nucleus of trophoblast in human placenta.

Panel D : anti-CD34 staining endothelium in human placenta.

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

The section was incubated in three rounds of staining : in the order of ab307087, ab218107 and ab315802 for 30 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.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling AP2 gamma/TFAP2C with ab218107 at 1/500 dilution followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on MCF7 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling AP2 gamma/TFAP2C with ab218107 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Almost all tumor cells show negative staining on moderately differentiated human breast cancer. The data is consistent with the literature (PMID : 18825690).

Counter stained with Hematoxylin.

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized SK-BR-3 (Human mammary gland adenocarcinoma cell line) cells labeling AP2 gamma/TFAP2C with ab218107 at 1/500 dilution followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on SK-BR-3 cell line.

The nuclear counterstain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling AP2 gamma/TFAP2C with ab218107 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Nuclear staining on human placenta 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) Ready to use.

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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed SK-BR-3 (Human mammary gland adenocarcinoma cell line) cells labeling AP2 gamma/TFAP2Cwith ab218107 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling AP2 gamma/TFAP2C with ab218107 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Nuclear staining on human normal breast 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) Ready to use.

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-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Immunohistochemical analysis of paraffin-embedded human seminoma tissue labeling AP2 gamma/TFAP2C with ab218107 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Nucleus staining on human seminoma 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) Ready to use.

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

• IP

Supplier Data

Immunoprecipitation - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

AP2 gamma/TFAP2C was immunoprecipitated from 0.35 mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab218107 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab218107 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : MCF7 whole cell lysate, 10 μg (Input).

Lane 2 : ab218107 IP in MCF7 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab2018107 in MCF7 whole cell lysate.

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (ab218107)

Predicted band size: 49 kDa

Observed band size: 49 kDa

false

• WB

Supplier Data

Western blot - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (AB218107)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1/2 : 30 seconds; Lane 3 : 10 seconds; Lane 4 : 3 minutes.

Lanes 1 - 2:

Western blot - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (ab218107) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-AP2 gamma/TFAP2C antibody [EPR20331] (ab218107) at 1/2000 dilution

Lane 1:

SK-BR-3 (Human mammary gland adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 2:

MDA-MB-435S (Human ductal carcinoma cell line) whole cell lysate at 10 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 4:

T-47D (Human ductal breast epithelial tumor cell line) 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: 49 kDa

Observed band size: 49 kDa

false