Anti-TCF7 antibody [EPR28579-5]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TCF7 antibody [EPR28579-5] (AB315392)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling TCF7 with ab315392 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human lung. The section was incubated with ab315392 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-TCF7 antibody [EPR28579-5] (AB315392)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling TCF7 with ab315392 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human skeletal muscle. The section was incubated with ab315392 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-TCF7 antibody [EPR28579-5] (AB315392)

Immunohistochemical analysis of paraffin-embedded Human Hodgkin's Lymphoma tissue labeling TCF7 with ab315392 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human Hodgkin's Lymphoma. The section was incubated with ab315392 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-TCF7 antibody [EPR28579-5] (AB315392)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling TCF7 with ab315392 at 1/2000 (0.245 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil (PMID : 12707037). The section was incubated with ab315392 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.

• mIHC

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Multiplex immunohistochemistry - Anti-TCF7 antibody [EPR28579-5] (AB315392)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human tonsil tissue staining TCF7 with ab315392 at a 1 : 2000 (0.26 ug/ml) dilution, ab237707 anti-CD3E used at 1 : 500 (1.01 ug/ml) dilution and ab237772 anti-CD19 used at a 1 : 5000 (0.21 ug/ml) dilution.

Panel A : merged staining of anti-TCF7 (magenta; Opal™690), anti-CD3E (green; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil.
Panel B : anti-TCF7 staining T lymphocytes of human tonsil.
Panel C : anti-CD3E staining T lymphocytes of human tonsil.
Panel D : anti-CD19 staining B lymphocytes of human tonsil.

The section was incubated in three rounds of staining : in the order of ab315392, ab237707, and ab237772 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.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TCF7 antibody [EPR28579-5] (AB315392)

Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized MOLT-4 (human lymphoblastic leukemia T lymphoblast) cells labelling TCF7 with ab315392 at 1/50 (9.8 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing nuclear staining in MOLT-4 cell line. Low expression : LNCaP (PMID : 25436980). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). 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 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-TCF7 antibody [EPR28579-5] (AB315392)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized LNCaP (human prostate carcinoma epithelial cell, Left) / MOLT-4 (human lymphoblastic leukemia T lymphoblast, Right) cells labelling TCF7 with ab315392 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : LNCaP.

• IP

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Immunoprecipitation - Anti-TCF7 antibody [EPR28579-5] (AB315392)

TCF7 was immunoprecipitated from 0.35 mg MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate with ab315392 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315392 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-TCF7 antibody [EPR28579-5] (ab315392) at 1/1000 dilution

Lane 1:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate

Lane 2:

ab315392 at 1/30 IP in MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab315392 in MOLT-4 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 32s

• WB

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Western blot - Anti-TCF7 antibody [EPR28579-5] (AB315392)

Low expression : LNCaP (PMID25436980).

This antibody should detect all TCF1/TCF7 isoforms, including those lacking the amino-terminal β-catenin binding domain.

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

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

All lanes:

Western blot - Anti-TCF7 antibody [EPR28579-5] (ab315392) at 1/1000 dilution

Lane 1:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 2:

LNCaP (human prostate carcinoma epithelial cell) whole cell 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 1/100000 dilution

Observed band size: 28-54 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-TCF7 antibody [EPR28579-5] (AB315392)

This antibody should detect all TCF1/TCF7 isoforms, including those lacking the amino-terminal β-catenin binding domain.

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

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

All lanes:

Western blot - Anti-TCF7 antibody [EPR28579-5] (ab315392) at 1/1000 dilution

Lane 1:

Human thymus tissue lysate at 20 µg

Lane 2:

Human tonsil 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 1/100000 dilution

Observed band size: 28-54 kDa,36 kDa

false

Exposure time: 26s