Anti-PYK2 antibody [EP206Y]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PYK2 antibody [EP206Y] (AB81266)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling PYK2 with ab81266 at 1/2000 (0.445 ug/ml) dilution.

Positive staining on human cerebrum.

The sections were using heat mediated antigen retrieval with EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins, then labeling ab81266 at 1/2000 dilution for 30 mins at room temperature. Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PYK2 antibody [EP206Y] (AB81266)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling PYK2 with ab81266 at 1/2000 (0.445 ug/ml) dilution.

Positive staining on rat cerebrum.

The sections were using heat mediated antigen retrieval with EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins, then labeling ab81266 at 1/2000 dilution for 30 mins at room temperature. Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PYK2 antibody [EP206Y] (AB81266)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling PYK2 with ab81266 at 1/2000 (0.445 ug/ml) dilution.

Positive staining on mouse cerebrum.

The sections were using heat mediated antigen retrieval with EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins, then labeling ab81266 at 1/2000 dilution for 30 mins at room temperature. Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Lab

Western blot - Anti-PYK2 antibody [EP206Y] (AB81266)

Western blot : Rabbit Monoclonal[EP206Y] to PYK2 ab81266 staining at 1/2500 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 120 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in PTK2B knockout HCT 116 cell line (ab301190). We are unable to identify the non-specific bands at 75kDa and 40kDa.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PYK2 antibody [EP206Y] (ab81266) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

Western blot - Human PTK2B knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-ptk2b-knockout-hct116-cell-line-ab301190'>ab301190</a>) at 20 µg

Lane 3:

Wild-type A549 at 20 µg

Lane 4:

PTK2B knockout A549 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 116 kDa

Observed band size: 120 kDa

false

• WB

Lab

Western blot - Anti-PYK2 antibody [EP206Y] (AB81266)

Western blot : Rabbit Monoclonal [EP206Y] to PYK2 ab81266 staining at 1/2000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 116 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in PTK2B knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween 20 (TBS-T) before incubation with primary antibodies overnight at 4 C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PYK2 antibody [EP206Y] (ab81266) at 1/2000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

Western blot - Human PTK2B knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-ptk2b-knockout-hct116-cell-line-ab301190'>ab301190</a>) at 20 µg

Lane 3:

A549 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 116 kDa

Observed band size: 116 kDa

false

• WB

Lab

Western blot - Anti-PYK2 antibody [EP206Y] (AB81266)

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

All lanes:

Western blot - Anti-PYK2 antibody [EP206Y] (ab81266) at 1/5000 dilution

All lanes:

Human brain lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 116 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-PYK2 antibody [EP206Y] (AB81266)

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

All lanes:

Western blot - Anti-PYK2 antibody [EP206Y] (ab81266) at 1/2500 dilution

Lane 1:

Mouse brain lysate at 15 µg

Lane 2:

Rat brain lysate at 15 µg

Secondary

All lanes:

minimal cross-reactivity with human IgG) at 1/2000 dilution

Predicted band size: 116 kDa

false

Exposure time: 180s