Anti-PSAP antibody [EPR25650-11] (BSA and Azide free)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300469, the same antibody clone in a different buffer formulation.Immunohistochemical

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling PSAP with ab300469 at 1/5000 (0.294 ug/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used. Positive staining on mouse spleen (PMID : 9570738). The section was incubated with ab300469 for 10 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) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300469, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling PSAP with ab300469 at 1/5000 (0.294 ug/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used. Positive staining on rat cerebral cortex (PMID : 9570738). The section was incubated with ab300469 for 10 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) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300469, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebral corte tissue labeling PSAP with ab300469 at 1/5000 (0.294 ug/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used. Positive staining on mouse cerebral cortex. The section was incubated with ab300469 for 10 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) was used.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-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300469, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Hepa1-6 (mouse hepatoma epithelial cell) cells labelling PSAP with ab300469 at 1/50 (11.74 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 2 μg/mL dilution (Green). Confocal image showing cytoplasmic staining in Hepa1-6 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^®594) was used to counterstain tubulin at 1/200 2.5 μg/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 μg/mL dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300469, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cortex (fresh) tissue labeling PSAP with ab300469 at 1/500 (1.174 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green). Positive staining on mouse cortex is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 2 μg/mL dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300469, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C6 (rat glial tumor glial cell) cells labelling PSAP with ab300469 at 1/50 (11.74 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 2 μg/mL dilution (Green). Confocal image showing cytoplasmic staining in C6 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5 μg/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 μg/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300469, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cortex (fresh) tissue labeling PSAP with ab300469 at 1/100 (5.87 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green). Positive staining on rat cortex is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 2 μg/mL dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300469, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C6 (rat glial tumor glial cell) cells labelling PSAP with ab300469 at 1/500 dilution (0.1ug) (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). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300469, the same antibody clone in a different buffer formulation.

Intracelluar flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Hepa1-6 (mouse hepatoma epithelial cell) cells labelling PSAP with ab300469 at 1/500 dilution (0.1 μg) (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). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using ab300470, the same antibody clone in a different buffer formulation.

PSAP was immunoprecipitated from 0.35 mg Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate 10 µg with ab300469 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300469 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate 10 µg Lane 2 : ab300469 IP in Hepa1-6 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300469 in Hepa1-6 whole cell lysate. Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 32 seconds

Lane 1:

Immunoprecipitation - Anti-PSAP antibody [EPR25650-11] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25650-11-ab300469'>ab300469</a>) at 1/30 dilution

Lane 2:

Immunoprecipitation - Anti-PSAP antibody [EPR25650-11] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25650-11-ab300469'>ab300469</a>) at 1/1000 dilution

All lanes:

Hepa1-6 whole cell lysate at 10 µg

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

Lab

Western blot - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using 300469, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-PSAP antibody [EPR25650-11] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25650-11-ab300469'>ab300469</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control, whole cell lysate at 20 µg

Lane 2:

NIH/3T3 transfected with siRNA 1 specifically targeting Prosaposin, whole cell lysate at 20 µg

Lane 3:

NIH/3T3 transfected with siRNA 2 specifically targeting Prosaposin, whole cell lysate at 20 µg

Lane 4:

NIH/3T3 transfected with siRNA 3 specifically targeting Prosaposin, 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>)

Predicted band size: 58 kDa

Observed band size: 60-80 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using 300469, 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 : 28541286).

Exposure time : Lane 1-3 : 103 seconds Lane 4 : 59 seconds

All lanes:

Western blot - Anti-PSAP antibody [EPR25650-11] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25650-11-ab300469'>ab300469</a>) at 1/1000 dilution

Lane 1:

Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

Mouse cerebellum tissue lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Lane 4:

Rat testis 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

Predicted band size: 58 kDa

Observed band size: 12 kDa,60 kDa,80 kDa

false

• WB

Lab

Western blot - Anti-PSAP antibody [EPR25650-11] (BSA and Azide free) (AB300470)

This data was developed using 300469, 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 : 28541286).

Exposure time : Lane 1 : 3 minutes Lane 2 : 70 seconds

All lanes:

Western blot - Anti-PSAP antibody [EPR25650-11] (<a href='/en-us/products/primary-antibodies/psap-antibody-epr25650-11-ab300469'>ab300469</a>) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), 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

Predicted band size: 58 kDa

Observed band size: 12 kDa,60 kDa,80 kDa

false