Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free

8 product images

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  Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free (ab236439)

  Immunohistochemistry (Frozen) analysis of mouse skin tissue section labeling Cytokeratin 14 with purified Anti-Cytokeratin 14 antibody [SP53] ab119695 at 1/500 (3.8 μg/ml). Sections were fixed in 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Antigen retrieval was heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 14 antibody [SP53] ab119695).

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free (ab236439)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Cytokeratin 14 antibody [SP53] ab119695). Anti-Cytokeratin 14 antibody [SP53] ab119695 staining KRT14 in wild-type A431 cells (top panel) and KRT14 knockout A431 cells (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-Cytokeratin 14 antibody [SP53] ab119695 at 5μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

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  Western blot - Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free (ab236439)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Cytokeratin 14 antibody [SP53] ab119695).
  

  Lanes 1 - 4: Merged signal (red and green). Green - Anti-Cytokeratin 14 antibody [SP53] ab119695 observed at 52 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

  Anti-Cytokeratin 14 antibody [SP53] ab119695 was shown to react with KRT14 in A431 wild-type cells in Western blot. Loss of signal was observed when KRT14 knockout sample was used. A431 wild-type and KRT14 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween^®) before incubation with Anti-Cytokeratin 14 antibody [SP53] ab119695 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 93 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Cytokeratin 14 antibody [SP53] (Anti-Cytokeratin 14 antibody [SP53] ab119695) at 1/93 dilution

  Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

  Lane 2: KRT14 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

  Lane 3: Human skin whole tissue lysate at 20 µg

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

  Performed under reducing conditions.

  Predicted band size: 52 kDa

  Observed band size: 52 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free (ab236439)

  Immunocytochemistry/ Immunofluorescence analysis of A431 (human epidermoid carcinoma epithelial cell) cells labeling Cytokeratin 14 with purified Anti-Cytokeratin 14 antibody [SP53] ab119695 at 1/2000 (0.95 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 14 antibody [SP53] ab119695).

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  Flow Cytometry (Intracellular) - Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free (ab236439)

  Intracellular Flow Cytometry analysis of A431 (human epidermoid carcinoma) labeling Cytokeratin 14 with purified Anti-Cytokeratin 14 antibody [SP53] ab119695 at 1/1900 dilution (1.01μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as a secondary antibody. Isotypecontrol - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black). Unlableled control - Unlabelled cells (blue).
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 14 antibody [SP53] ab119695).

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  Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free (ab236439)

  Immunohistochemistry (Frozen) analysis of rat skin tissue section labeling Cytokeratin 14 with purified Anti-Cytokeratin 14 antibody [SP53] ab119695 at 1/500 (3.8 μg/ml). Sections were fixed in 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Antigen retrieval was heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cytokeratin 14 antibody [SP53] ab119695).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free (ab236439)

  Formalin-fixed, Paraffin-embedded Human prostate tissue stained for Cytokeratin 14 uisng ab119700 in Immunohistochemical analysis.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab119700).

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  Multiplex immunohistochemistry - Anti-Cytokeratin 14 antibody [SP53] - BSA and Azide free (ab236439)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast labelling Cytokeratin 18 with Anti-Cytokeratin 18 antibody [SP69] ab93741 at 1/200 dilution (1.02 µg/mL) (B), Collagen VI with Anti-Collagen VI antibody [EPR17072] - BSA and Azide free ab271938 at 1/500 dilution (2.084 μg/ml) (C) and Cytokeratin 14 with ab236439 at 1/2000 dilution ( 0.519 μg/ml) (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
  
  Panel A: merged staining of anti-Cytokeratin 14 (magenta; Opal™690), anti-Cytokeratin 18 (green; Opal™520) and anti-Collagen VI (red; Opal™570) on human breast.
  Panel B: anti-Cytokeratin 18 stained on luminal epithelial cells.
  Panel C: anti-Collagen VI stained on stroma.
  Panel D: anti-Cytokeratin 14 stained on myoepithelial cells.
  
  The section was incubated in three rounds of staining: in the order of ab236439 for 30 mins, Anti-Cytokeratin 18 antibody [SP69] ab93741 for 10 mins, and Anti-Collagen VI antibody [EPR17072] - BSA and Azide free ab271938 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.