Anti-Hes1 antibody [EPR4226]

6 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-Hes1 antibody [EPR4226] (ab108937)

  Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% TritonX-100 permeabilised SH-SY5Y (Human neuroblastoma epithelial cell) labelling Hes1 with ab108937 at 1/1000 dilution, followed by AlexaFluor^® 488 Goat anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in SH-SY5Y cell line is observed. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) was used to counterstain alpha Tubulin at 1/200 dilution (red). The Nuclear counterstain was DAPI (blue).
  

  Secondary antibody only control: Secondary antibody is AlexaFluor® 488 Goat anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.

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  Western blot - Anti-Hes1 antibody [EPR4226] (ab108937)

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

  All lanes: Western blot - Anti-Hes1 antibody [EPR4226] (ab108937) at 1/1000 dilution

  Lane 1: SH-SY5Y (Human bone marrow epithelial) whole cell lysate at 20 µg

  Lane 2: MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

  Lane 3: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 4: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 5: K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

  Lane 6: HUVEC (Human umbilical vein endothelial cell) whole cell lysate at 20 µg

  Lane 7: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 8: Human brain tissue lysate at 20 µg

  Lane 9: Human liver tissue lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/20000 dilution

  Predicted band size: 30 kDa

  Observed band size: 30 kDa

  Exposure time: 120s

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hes1 antibody [EPR4226] (ab108937)

  Primary ab concentration dilution: 1:200, (0.5ug/ml)
  

  Secondary ab: ImmunoHistoprobe (Ready to use) HRP Polymer for Rabbit IgG

  Secondary ab concentration: Prediluted

  Tissue: Human brain

  Fixative: Paraffin-embedded sections

  Counter stain: Hematoxylin

  Antigen retrieval: Perform heat mediated antigen retrieval using Tris/EDTA Buffer, PH9

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  Flow Cytometry (Intracellular) - Anti-Hes1 antibody [EPR4226] (ab108937)

  Overlay histogram showing SH-SY5Y cells stained with unpurifiedab108937 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab108937, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hes1 antibody [EPR4226] (ab108937)

  Immunohistochemical staining of Hes1 in paraffin-embedded human placenta tissue with unpurified ab108937 at 1/250 dilution.
  

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hes1 antibody [EPR4226] (ab108937)

  Immunohistochemical staining of paraffin embedded human brain with unpurified ab108937 at a working dilution of 1 in 90. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.