Anti-HEXA antibody [EPR26394-74]

9 product images

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  Immunoprecipitation - Anti-HEXA antibody [EPR26394-74] (ab300447)

  HEXA was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 ug with ab300447 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300447 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: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 ug

  Lane 2: abAB300447 IP in RAW264.7 whole cell lysate

  Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab300447 in RAW264.7 whole cell lysate

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

  Exposure time: 3.25 seconds

  All lanes: Immunoprecipitation - Anti-HEXA antibody [EPR26394-74] (ab300447) at 1/1000 dilution

  All lanes: RAW264.7 whole cell 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

  Exposure time: 3.25s

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  Flow Cytometry (Intracellular) - Anti-HEXA antibody [EPR26394-74] (ab300447)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-12 (Rat adrenal gland pheochromocytoma) cells labelling HEXA with ab300447 at 1/500 dilution (0.1ug) (Red) (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.

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  Flow Cytometry (Intracellular) - Anti-HEXA antibody [EPR26394-74] (ab300447)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling HEXA with ab300447 at 1/500 dilution (0.1ug) (Red) (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.

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  Immunocytochemistry/ Immunofluorescence - Anti-HEXA antibody [EPR26394-74] (ab300447)

  Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma small irregularly shaped cells) cells labelling HEXA with ab300447 at 1/50 (11.54 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 PC-12 cell line is observed. 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 2.5ug/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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  Immunocytochemistry/ Immunofluorescence - Anti-HEXA antibody [EPR26394-74] (ab300447)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling HEXA with ab300447 at 1/50 (11.54 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 RAW 264.7 cell line is observed. 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 2.5ug/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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  Western blot - Anti-HEXA antibody [EPR26394-74] (ab300447)

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

  All lanes: Western blot - Anti-HEXA antibody [EPR26394-74] (ab300447) at 1/1000 dilution

  Lane 1: Mouse testis tissue lysate

  Lane 2: Mouse heart tissue lysate

  Lane 3: Mouse liver tissue lysate

  Lane 4: Rat testis tissue lysate

  Lane 5: Rat liver tissue lysate

  Lane 6: Rat heart tissue lysate

  Secondary

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

  Performed under reducing conditions.

  Observed band size: 61 kDa, 54 kDa

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  Western blot - Anti-HEXA antibody [EPR26394-74] (ab300447)

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

  All lanes: Western blot - Anti-HEXA antibody [EPR26394-74] (ab300447) at 1/1000 dilution

  Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

  Lane 2: C2C12 (mouse myoblasts myoblast) whole cell lysate

  Lane 3: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

  Lane 4: C6 (rat glial tumor glial cell) whole cell lysate

  Lane 5: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Performed under reducing conditions.

  Observed band size: 61 kDa, 54 kDa

  Exposure time: 26s

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HEXA antibody [EPR26394-74] (ab300447)

  Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling HEXA with ab300447 at 1/2000 (0.289 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on rat testis. The section was incubated with ab300447 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) was used.Heat mediated antigen retrieval 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-HEXA antibody [EPR26394-74] (ab300447)

  Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling HEXA with ab300447 at 1/2000 (0.289 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse colon. The section was incubated with ab300447 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) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins