Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling EGFR with purified ab40815 at 1/500 dilution (1.75 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

Immunofluorescent staining of untreated (C) and Phosphatase- treated (D) A431 cells using ab40815 (unpurified).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) treated with 200 ng/ml EGF for 15 minutes cells labeling EGFR with purified ab40815 at 1/800 dilution (1μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). Untreated A431 cells (Green).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

WD-PBEC cultures were infected with RSV clinical isolate BT2a and stained for EGFR (red) and RSV F (RSV F, green) expression.

For WD-PBECs, pediatric bronchial epithelial cells (PBEC) were obtained, via written parental consent, from bronchial brushings of children undergoing elective surgery at the Royal Belfast Hospital for Sick Children, and the procedures were approved by the Office of Research Ethics Committees Northern Ireland. PBEC were expanded in collagen-coated flasks using airway epithelial cell media and supplements (Lonza), then seeded onto transwell inserts (Corning), and then air-liquid interface (ALI) cultures were initiated and maintained 21 days in order to establish well-differentiated (WD)-PBECs. Paraformaldehyde-fixed and permeabilized WD-PBEC were stained for RSV F protein expression and were stained with anti-phospho-(p)-EGFR (Abcam, ab40815). WD-PBEC cultures were infected with RSV subgroup A clinical isolate BT2a. Fluorescent images were obtained with a SP5 confocal DMI 6000 inverted microscope (Leica).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

Clone EP774Y (ab182618) has been successfully conjugated by Abcam. This image was generated using Anti-EGFR (phospho Y1068) antibody [EP774Y] (Alexa Fluor® 488). Please refer to ab205827 for protocol details.

ab205827 staining EGFR (phospho Y1068) in A431 cells +/-EGF (100ng/ml, 5min). The cells were fixed with 4% formaldehyde (10 min), 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 ab205827 at 1 : 100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

Clone EP774Y (ab182618) has been successfully conjugated by Abcam. This image was generated using Anti-EGFR (phospho Y1068) antibody [EP774Y] (Alexa Fluor® 647). Please refer to ab205828 for protocol details.

ab205828 staining EGFR (phospho Y1092) in A431 cells +/-EGF (100ng/ml, 5min). The cells were fixed with 4% formaldehyde (10 min), 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 ab205828 at 1 : 100 dilution (shown in red) and 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).

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 10 minutes cells labeling EGFR with purified ab40815 at 1/500 dilution (1.8 μg/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) at 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

• IHC-P

AbReview55755****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

Formaldehyde-fixed, paraffin-embedded human prostate cancer tissue stained for EGFR (phospho Y1068) using ab40815 (unpurified) at 1/200 dilution in immunohistochemical analysis, followed by Goat anti Rabbit IgG (Biotin).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

This image is courtesy of an anonymous Abreview.

• WB

Supplier Data

Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

***Blocking and dilution buffer : *** 5% NFDM/TBST. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

Lanes 1 - 3:

Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] (<a href='/en-us/products/primary-antibodies/egfr-phospho-y1068-antibody-ep774y-ab40815'>ab40815</a>) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (ab182618) at 1/1000 dilution

Lane 1:

C2C12 (Mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 2:

C2C12 (Mouse myoblasts myoblast) treated with 100 ng/ml EGF for 24 hours whole cell lysate at 20 µg

Lane 3:

C2C12 (Mouse myoblasts myoblast) treated with 100 ng/ml EGF for 24hours whole cell lysate, then the membrane was incubated with phosphatase 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

false

Exposure time: 30s

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

Immunohistochemical staining of untreated (A) and Phosphatase- treated (B) paraffin-embedded breast adenocarcinoma tissue using ab40815 (unpurified).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

ab40815 (unpurified) showing positive staining in Cervical carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

ab40815 (unpurified) showing positive staining in Glioma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

ab40815 (unpurified) showing positive staining in Papillary carcinoma of thyroid gland tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).

• OI-RD Scanning

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OI-RD Scanning - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• Dot

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Dot Blot - Anti-EGFR (phospho Y1068) antibody [EP774Y] - BSA and Azide free (AB182618)

Dot blot analysis of EGFR (pY1068) peptide (Lane 1), SMAD5 (unmodified) peptide labelling EGFR (pY1068) with ab40815 (unpurified) at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40815).