Anti-PSAP antibody [EPR25649-20] - BSA and Azide free

• IHC

Supplier Data

Immunohistochemistry - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was produced using ab308122, the same antibody clone but in a different formulation. Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling PSAP with ab308122 at 1/2000 followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive cytoplasmic staining on human cerebrum. The section was incubated with ab308122 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

Supplier Data

Immunohistochemistry - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was produced using ab308122, the same antibody clone but in a different formulation. Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling PSAP with ab308122 at 1/2000 followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive cytoplasmic staining on human spleen. The section was incubated with ab308122 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was produced using ab308122, the same antibody clone but in a different formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-3 (human prostate adenocarcinoma epithelial cell) cells labelling PSAP with ab308122 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in PC-3 cell line. 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 dilution (2.5 ug/ml) (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 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was developed using ab308122, the same antibody clone in a different buffer formulation. PSAP was immunoprecipitated from 0.35 mg PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate with ab308122 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308122 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : PC-3 whole cell lysate Lane 2 : ab308122 IP in PC-3 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308122 in PC-3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 8 seconds.

All lanes:

Immunoprecipitation - Anti-PSAP antibody [EPR25649-20] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25649-20-ab308122'>ab308122</a>) at 1/30 dilution

Lane 1:

PC-3 whole cell lysate at 20 µg

Lane 2:

PC-3 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

Observed band size: 70 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was developed using ab308122, the same antibody clone in a different buffer formulation.

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

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.

The band below 70 kDa may be caused by degradation.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PSAP antibody [EPR25649-20] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25649-20-ab308122'>ab308122</a>) at 1/1000 dilution

All lanes:

Human cerebellum 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: 70 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was developed using ab308122, the same antibody clone in a different buffer formulation.

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

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.

Lysates were freshly made and used immediately to minimize protein degradation.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 15743835) (PMID : 15353235).

Exposure time :

Lane 1 : 180 seconds

Lane 2 : 15 seconds

All lanes:

Western blot - Anti-PSAP antibody [EPR25649-20] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25649-20-ab308122'>ab308122</a>) at 1/1000 dilution

Lane 1:

HeLa whole cell lysate at 20 µg

Lane 2:

PC-3 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: 70 kDa

true

• WB

Supplier Data

Western blot - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was developed using ab308122, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 15743835) (PMID : 15353235). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-PSAP antibody [EPR25649-20] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25649-20-ab308122'>ab308122</a>) at 1/1000 dilution

Lane 1:

HeLa transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting PSAP 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: 70 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was developed using ab308122, the same antibody clone in a different buffer formulation.

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 15743835) (PMID : 15353235).

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.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PSAP antibody [EPR25649-20] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25649-20-ab308122'>ab308122</a>) at 1/1000 dilution

Lane 1:

293T transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically targeting PSAP 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 1/20000 dilution

Observed band size: 70 kDa

true

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was developed using ab308122, the same antibody clone in a different buffer formulation.

ab308122 was shown to react with PSAP in wild-type HeLa cells in Western blot with loss of signal observed in PSAP knockout cell line ab265989. Wild-type HeLa and PSAP knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab308122 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 μg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-PSAP antibody [EPR25649-20] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25649-20-ab308122'>ab308122</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa lysate at 10 µg

Lane 2:

PSAP knock-out HeLa lysate at 10 µg

false

• IP

Supplier Data

Immunoprecipitation - Anti-PSAP antibody [EPR25649-20] - BSA and Azide free (AB308123)

This data was developed using ab308122, the same antibody clone in a different buffer formulation.

Immunoprecipitation of PSAP in HeLa cells. Lysates were prepared and immunoprecipitation was performed using 2 μg of ab308122 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

HeLa cells

false