Anti-Lewis a,b carbohydrate antibody [HEA164]

7 product images

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  Flow Cytometry (Intracellular) - Anti-Lewis a,b carbohydrate antibody [HEA164] (ab322960)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell, Left) / BxPC-3 (human pancreas adenocarcinoma epithelial cell, Right) cells labeling Lewis a,b carbohydrate with ab322960 at 1/100 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

  Negative control: HepG2

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lewis a,b carbohydrate antibody [HEA164] (ab322960)

  Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Lewis a,b carbohydrate with ab322960 at 1/10000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Low expression: no staining on human liver.
  
  The section was incubated with ab322960 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).

  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-Lewis a,b carbohydrate antibody [HEA164] (ab322960)

  Immunohistochemical analysis of paraffin-embedded Human pancreas cancer tissue labeling Lewis a,b carbohydrate with ab322960 at 1/10000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human pancreas cancer.
  
  The section was incubated with ab322960 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).

  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-Lewis a,b carbohydrate antibody [HEA164] (ab322960)

  Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling Lewis a,b carbohydrate with ab322960 at 1/10000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human colon cancer.
  
  The section was incubated with ab322960 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).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lewis a,b carbohydrate antibody [HEA164] (ab322960)

  Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Lewis a,b carbohydrate with ab322960 at 1/10000 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human colon.
  
  The section was incubated with ab322960 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).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

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  Immunocytochemistry/ Immunofluorescence - Anti-Lewis a,b carbohydrate antibody [HEA164] (ab322960)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized BxPC-3 (human pancreas adenocarcinoma epithelial cell) cells labelling Lewis a,b carbohydrate with ab322960 at 1/2000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green).

  Confocal image showing cytoplasmic and membranous staining in BxPC-3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
  
  Negative control: HepG2.
  
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

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  Western blot - Anti-Lewis a,b carbohydrate antibody [HEA164] (ab322960)

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

  Negative control: HepG2
  
  Low expression tissue: human liver

  The identity of the lower MW band at approximately 150 kDa (in lane 3) is unknown.

  Exposure time: Lane 1: 8 seconds; Lane2-4: 59 seconds

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  All lanes: Western blot - Anti-Lewis a,b carbohydrate antibody [HEA164] (ab322960) at 1/1000 dilution

  Lane 1: BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: Human pancreas cancer tissue lysate at 20 µg

  Lane 4: Human liver tissue lysate at 20 µg

  Secondary

  All lanes: Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

  Observed band size: 250 kDa, 36 kDa