Anti-Cytokeratin 4 antibody [EP1599Y]

5 product images

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)

  Immunohistochemical staining of paraffin embedded human cervical carcinoma tissue using ab51599 (1/100). Goat anti-rabbit HRP (1/2000) used for secondary staining.
  

  Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)

  ICC/IF image of ab51599 stained A431 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab51599 at 1/100 dilution overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43μM for 1hour at room temperature.

• 

  Western blot - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)

  All lanes: Western blot - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599) at 1/1000 dilution

  All lanes: A431 cell lysate at 10 µg

  Secondary

  All lanes: Goat anti-rabbit HRP at 1/2000 dilution

  Predicted band size: 57 kDa

  Observed band size: 59 kDa

• 

  Immunoprecipitation - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)

  Purified ab51599 at 1/50 dilution (2μg) immunoprecipitating Cytokeratin 4 in HaCaT whole cell lysate.
  Lane 1 (input): HaCaT (Human skin keratinocyte) whole cell lysate 10μg
  Lane 2 (+): ab51599 + HaCaT whole cell lysate.
  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab51599 in HaCaT whole cell lysate.
  VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/1000 dilution) was used for Western blotting.
  Blocking Buffer and concentration: 5% NFDM/TBST.
  Diluting buffer and concentration: 5% NFDM/TBST.
  Observed band size: 59 kDa

  All lanes: Immunoprecipitation - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)

  Predicted band size: 57 kDa

• 

  Flow Cytometry (Intracellular) - Anti-Cytokeratin 4 antibody [EP1599Y] (ab51599)

  Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 4 with purified ab51599 at 1/100 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue).