Anti-HMGCS2 antibody [EPR26978-39]

8 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGCS2 antibody [EPR26978-39] (ab305227)

  Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HMGCS2 with ab305227 at 1/2000 (0.237 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on human colon (PMID: 35422042, PMID: 31355161). The section was incubated with ab305227 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution.
  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

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  Dot Blot - Anti-HMGCS2 antibody [EPR26978-39] (ab305227)

  Dot blot analysis of HMGCS2 using ab305227 at 1:1000 (0.473 μg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.
  
  Lane 1: His-tagged human HMGCS1 recombinant protein 10 ng
  Lane 2: MYC/DDK tagged human HMGCS2 recombinant protein 10 ng
  
  Exposure time: 180 seconds
  
  Blocking and diluting buffer and concentration: 5% NFDM/TBST

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  Immunocytochemistry/ Immunofluorescence - Anti-HMGCS2 antibody [EPR26978-39] (ab305227)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Caco-2 (human colorectal adenocarcinoma epithelial cell) cells labelling HMGCS2 with ab305227 at 1/500 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 mitochondrial staining in Caco-2 cells. Negative control: HCT 116 (PMID: 28468827). The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
  Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker ab33985 Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/1000 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 dilution.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGCS2 antibody [EPR26978-39] (ab305227)

  Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling HMGCS2 with ab305227 at 1/2000 (0.237 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Negative control: no staining on human cerebrum (PMID: 7893153, PMID: 22523078, PMID: 23943793, PMID: 24391449). The section was incubated with ab305227 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution.
  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-HMGCS2 antibody [EPR26978-39] (ab305227)

  Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HMGCS2 with ab305227 at 1/2000 (0.237 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on human liver (PMID: 31779269). The section was incubated with ab305227 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution.
  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

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  Immunoprecipitation - Anti-HMGCS2 antibody [EPR26978-39] (ab305227)

  HMGCS2 was immunoprecipitated from 0.35 mg Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate 10 μg with ab305227 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305227 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: Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate 10 μg
  
  Lane 2: ab305227 IP in Caco-2 whole cell lysate
  
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab305227 in Caco-2 whole cell lysate
  
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  
  Exposure time: 180 seconds

  All lanes: Immunoprecipitation - Anti-HMGCS2 antibody [EPR26978-39] (ab305227) at 1/30 dilution

  All lanes: Caco-2 (human colorectal adenocarcinoma epithelial cell)

  Secondary

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

  Observed band size: 50 kDa

  Exposure time: 180s

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  Western blot - Anti-HMGCS2 antibody [EPR26978-39] (ab305227)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  Negative control: HCT 116 (PMID: 28468827)
  Exposure time:
  lanes 1-2: 6 seconds
  lanes 3-5: 180 seconds

  All lanes: Western blot - Anti-HMGCS2 antibody [EPR26978-39] (ab305227) at 1/1000 dilution

  Lane 1: Human liver tissue lysate at 20 µg

  Lane 2: Human colon tissue lysate at 20 µg

  Lane 3: COLO 205 (human colon adenocarcinoma tumor ascitic epithelial cell), whole cell lysate at 20 µg

  Lane 4: HCT 116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

  Lane 5: Caco-2 (human colorectal adenocarcinoma epithelial cell), 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: 50 kDa

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  Flow Cytometry (Intracellular) - Anti-HMGCS2 antibody [EPR26978-39] (ab305227)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HCT116 (human colorectal carcinoma epithelial cell, Left) / Caco-2 (human colorectal adenocarcinoma epithelial cell, Right) cells labelling HMGCS2 with ab305227 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: HCT116 (PMID: 28468827).