Anti-SPTLC1 antibody [EPR26226-136]

9 product images

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  Western blot - Anti-SPTLC1 antibody [EPR26226-136] (ab307552)

  SPTLC1 Western blot staining using rabbit Anti-SPTLC1 antibody

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

  Compared with ab307552, Anti-SPTLC1 antibody [EPR26226-221] ab315158 has higher sensitivity. We recommend Anti-SPTLC1 antibody [EPR26226-221] ab315158 as an alternative.

  Lanes 1 - 4: Western blot - Anti-SPTLC1 antibody [EPR26226-136] (ab307552) at 1/1000 dilution

  Lanes 5 - 8: Western blot - Anti-SPTLC1 antibody [EPR26226-221] (Anti-SPTLC1 antibody [EPR26226-221] ab315158) at 1/1000 dilution

  Lanes 1 and 5: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lanes 2 and 6: 293T (human embryonic kidney epithelial cell) whole cell lysate

  Lanes 3 and 7: HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

  Lanes 4 and 8: Human small intestine tissue lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Observed band size: 54 kDa

  Exposure time: 60s

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  Immunoprecipitation - Anti-SPTLC1 antibody [EPR26226-136] (ab307552)

  SPTLC1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 10 µg with ab307552 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307552 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: HeLa whole cell lysate

  Lane 2: ab307552 at 1/30 IP in HeLa whole cell lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab307552 in HeLa whole cell lysate

  Blocking and dilution 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.

  All lanes: Immunoprecipitation - Anti-SPTLC1 antibody [EPR26226-136] (ab307552) at 1/1000 dilution

  Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) at 10 µg

  Lane 2: ab307552 at 1/30 IP in HeLa whole cell lysate

  Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 in HeLa whole cell lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Developed using the ECL technique.

  Observed band size: 53 kDa

  Exposure time: 180s

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  Western blot - Anti-SPTLC1 antibody [EPR26226-136] (ab307552)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  This antibody does not cross-react with human SPTLC2 and SPTLC3.
  
  Exposure time: 37 seconds.

  All lanes: Western blot - Anti-SPTLC1 antibody [EPR26226-136] (ab307552) at 1/1000 dilution

  Lane 1: His-tagged human SPTLC1 recombinant protein fragment at 10 ng

  Lane 2: His-tagged human SPTLC2 recombinant protein fragment at 10 ng

  Lane 3: His-tagged human SPTLC3 recombinant protein fragment at 10 ng

  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: 46 kDa

  Exposure time: 37s

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  Western blot - Anti-SPTLC1 antibody [EPR26226-136] (ab307552)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  Negative control: human skeletal muscle (PMID:17023427).
  
  Exposure time: 48 seconds.

  All lanes: Western blot - Anti-SPTLC1 antibody [EPR26226-136] (ab307552) at 1/1000 dilution

  Lane 1: Human placenta tissue lysate at 20 µg

  Lane 2: Human skeletal muscle tissue lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Observed band size: 53 kDa

  Exposure time: 48s

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPTLC1 antibody [EPR26226-136] (ab307552)

  Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling SPTLC1 with ab307552 at 1/200 dilution (2.44 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
  Negative control: No staining on human skeletal muscle.
  The section was incubated with ab307552 at 4°C overnight. Counterstained with Hematoxylin.
  
  Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer.
  Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPTLC1 antibody [EPR26226-136] (ab307552)

  Immunohistochemical analysis of paraffin-embedded human colon tissue labeling SPTLC1 with ab307552 at 1/200 dilution (2.44 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
  Positive staining on human colon (PMID: 12704216).
  The section was incubated with ab307552 at 4°C overnight. Counterstained with Hematoxylin.
  
  Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer.
  Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPTLC1 antibody [EPR26226-136] (ab307552)

  Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling SPTLC1 with ab307552 at 1/200 dilution (2.44 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
  Positive staining on human cerebrum.
  The section was incubated with ab307552 at 4°C overnight. Counterstained with Hematoxylin.
  
  Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer.
  Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPTLC1 antibody [EPR26226-136] (ab307552)

  Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling SPTLC1 with ab307552 at 1/200 dilution (2.44 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
  Positive staining on human placenta.
  The section was incubated with ab307552 at 4°C overnight. Counterstained with Hematoxylin.
  
  Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer.
  Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

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  Flow Cytometry (Intracellular) - Anti-SPTLC1 antibody [EPR26226-136] (ab307552)

  Flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling SPTLC1 with ab307552 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabeled 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 1/2000 dilution was used as the secondary antibody.