Anti-PSPH antibody [EPR26995-88A]

• IHC

Supplier Data

Immunohistochemistry - Anti-PSPH antibody [EPR26995-88A] (AB306577)

Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling Phosphoserine phosphatase with ab306577 at 1/500 (1.078 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Negative control : no staining in human skeletal muscle. The section was incubated with ab306577 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 ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC

Supplier Data

Immunohistochemistry - Anti-PSPH antibody [EPR26995-88A] (AB306577)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Phosphoserine phosphatase with ab306577 at 1/500 (1.078 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining in human kidney. The section was incubated with ab306577 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 ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC

Supplier Data

Immunohistochemistry - Anti-PSPH antibody [EPR26995-88A] (AB306577)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Phosphoserine phosphatase with ab306577 at 1/500 (1.078 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining in human colon. The section was incubated with ab306577 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 ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-PSPH antibody [EPR26995-88A] (AB306577)

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

Low expression : MCF7 (PMID : 32783398) and A549 (PMID : 30874300).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-PSPH antibody [EPR26995-88A] (ab306577) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

A549 (human lung 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: 25 kDa

true

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-PSPH antibody [EPR26995-88A] (AB306577)

Blocking and diluting buffer and concentration : 5% NFDM/TBST   Exposure time : Lane 1 : 180 seconds Lane 2 : 15 seconds

All lanes:

Western blot - Anti-PSPH antibody [EPR26995-88A] (ab306577) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate 20 μg

Lane 2:

Human kidney tissue lysate 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: 25 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-PSPH antibody [EPR26995-88A] (AB306577)

Western blot : Anti-PSPH antibody [EPR26995-88A] (ab306577) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab306577 was shown to bind specifically to PSPH. A band was observed at 25 kDa in wild-type A549 cell lysates with no signal observed at this size in PSPH knockout cell line. To generate this image, wild-type and PSPH knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-PSPH antibody [EPR26995-88A] (ab306577) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

PSPH knockout A549 cell lysate at 20 µg

Lane 3:

Caco-2 cell lysate at 20 µg

Lane 4:

A549 Nuclear cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 25 kDa

false