Anti-Stra8 antibody [EPR27083-56]

11 product images

• 

  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 (Anti-GAPDH antibody [EPR16891] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Developed using the ECL technique.

  Observed band size: 55 kDa

  Exposure time: 180s

• 

  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.

• 

  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.

• 

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• 

  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 (Anti-GAPDH antibody [EPR16891] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 55 kDa

  Exposure time: 180s

• 

  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.

• 

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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). Alexa Fluor® 647 Anti-Myc tag antibody [9E10] 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• 

  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.

• 

  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.

• 

  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)(VeriBlot for IP Detection Reagent (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Observed band size: 55 kDa

  Exposure time: 180s

• 

  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 (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 55 kDa

  Exposure time: 92s