Anti-Stra8 antibody [EPR27083-56]

• IHC

Supplier Data

Immunohistochemistry - Anti-Stra8 antibody [EPR27083-56] (AB308124)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Stra8 with ab308124 at 1/2000 followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on rat testis. The section was incubated with ab308124 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-Stra8 antibody [EPR27083-56] (AB308124)

Immunohistochemical analysis of paraffin-embedded transfected cells labeling Stra8 with ab308124 at 1/2000 followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on HEK-293T transfected with a His-tagged mouse STRA8 construct (image A). No staining on HEK-293T transfected with empty plasmid (image B). The section was incubated with ab308124 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-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Stra8 antibody [EPR27083-56] (AB308124)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Stra8 with ab308124 at 1/2000 followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Negative control : no staining on rat spleen. The section was incubated with ab308124 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC

Supplier Data

Immunohistochemistry - Anti-Stra8 antibody [EPR27083-56] (AB308124)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Stra8 with ab308124 at 1/2000 followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on mouse testis (PMID : 8896602) (PMID : 32032549). The section was incubated with ab308124 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

• IHC

Supplier Data

Immunohistochemistry - Anti-Stra8 antibody [EPR27083-56] (AB308124)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Stra8 with ab308124 at 1/2000 followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection). Negative control : no staining on mouse spleen. The section was incubated with ab308124 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Stra8 antibody [EPR27083-56] (AB308124)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labelling Stra8 with ab308124 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 nuclear staining in F9 cells treated with Retinoic acid (1µM ) for 24 hours (PMID : 8896602). 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Stra8 antibody [EPR27083-56] (AB308124)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK293T (human embryonic kidney epithelial cell) cells labelling Stra8 with ab308124 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 nuclear staining in HEK-293T cells transfected with a mouse STRA8 expression vector containing a myc tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab223895 Anti-Myc tag Mouse monoclonal antibody (Alexa Fluor® 647) was used to counterstain tubulin at 1/200 dilution (10 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-Stra8 antibody [EPR27083-56] (AB308124)

This data was developed using ab308124, the same antibody clone in a different buffer formulation. Stra8 was immunoprecipitated from 0.35 mg mouse testis tissue lysate with ab308124 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308124 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse testis tissue lysate Lane 2 : ab308124 IP in mouse testis tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308124 in mouse testis tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-Stra8 antibody [EPR27083-56] (ab308124) at 1/30 dilution

All lanes:

Mouse testis tissue 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

Observed band size: 55 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Stra8 antibody [EPR27083-56] (AB308124)

This data was developed using ab308124, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. The expression of STRA8 is upregulated in response to 1 µM retinoic acid for 24 hours treatment (PMID : 8896602). In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution. Exposure time : 92 seconds.

All lanes:

Western blot - Anti-Stra8 antibody [EPR27083-56] (ab308124) at 1/1000 dilution

Lane 1:

Untreated F9 whole cell lysate at 40 µg

Lane 2:

F9 treated with 1 µM retinoic acid for 24 hours 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: 55 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-Stra8 antibody [EPR27083-56] (AB308124)

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.

Negative control : spleen (PMID : 8896602).

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

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

All lanes:

Western blot - Anti-Stra8 antibody [EPR27083-56] (ab308124) at 1/1000 dilution

Lane 1:

Rat testis tissue lysate at 20 µg

Lane 2:

Rat spleen 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: 55 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Stra8 antibody [EPR27083-56] (AB308124)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : spleen (PMID : 8896602). Lysates were freshly made and used immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds.

All lanes:

Western blot - Anti-Stra8 antibody [EPR27083-56] (ab308124) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse spleen 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: 55 kDa

false

Exposure time: 180s