Anti-SPARCL1 antibody [EPR28135-54]

6 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-SPARCL1 antibody [EPR28135-54] (ab313638)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling SPARCL1 with ab313638 at 1/50 (9.9 ug/ml) 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 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in mouse astrocyte cells.Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody was used to counterstain tubulin at 1/50 10ug/ml dilution (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 2ug/ml dilution.

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  Immunoprecipitation - Anti-SPARCL1 antibody [EPR28135-54] (ab313638)

  SPARCL1 was immunoprecipitated from 0.35 mg Mouse cerebellum tissue lysate with ab313638 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313638 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 cerebellum tissue lysate
  Lane 2: ab313638 IP in Mouse cerebellum tissue lysate
  Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab313638 in mouse cerebellum tissue lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  Exposure time: 10 seconds

  All lanes: Immunoprecipitation - Anti-SPARCL1 antibody [EPR28135-54] (ab313638) at 1/30 dilution

  All lanes: Mouse cerebellum tissue lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

  Exposure time: 10s

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  Western blot - Anti-SPARCL1 antibody [EPR28135-54] (ab313638)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  The molecular weight observed is consistent with what has been described in the literature (PMID: 22173850).
  Low expression: kidney (PMID: 9199668; PMID: 32437418).
  In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
  Exposure time: 6 seconds

  All lanes: Western blot - Anti-SPARCL1 antibody [EPR28135-54] (ab313638) at 1/1000 dilution

  Lane 1: Mouse brain tissue lysate at 20 µg

  Lane 2: Mouse cerebellum tissue lysate at 20 µg

  Lane 3: Mouse hippocampus tissue lysate at 20 µg

  Lane 4: Mouse lung tissue lysate at 20 µg

  Lane 5: Mouse kidney 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 1/20000 dilution

  Observed band size: 130 kDa

  Exposure time: 6s

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  Flow Cytometry (Intracellular) - Anti-SPARCL1 antibody [EPR28135-54] (ab313638)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neural/glia cell cells labelling SPARCL1 with ab313638 at 1/50 dilution (1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARCL1 antibody [EPR28135-54] (ab313638)

  Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling SPARCL1 with ab313638 at 1/2000 (0.248 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on endothelium of mouse spleen (PMID: 9199668).The section was incubated with ab313638 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background 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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARCL1 antibody [EPR28135-54] (ab313638)

  Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling SPARCL1 with ab313638 at 1/2000 (0.248 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum (PMID: 18381651).The section was incubated with ab313638 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background 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