Anti-Mucin 5AC antibody [45M1]

12 product images

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  Image from Kikuchi M et al., BMC Gastroenterol. 2010 Jun 18;10:65. doi: 10.1186/1471-230X-10-65.; Fig 5.; 18 June 2010, BMC Gastroenterology 2010, 10:65

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mucin 5AC antibody [45M1] (ab3649)

  This image was generated from the hybridoma version of the product.
  

  Immunohistochemical analysis of rat stomach tissue, staining Mucin 5AC with ab3649.
  
  Antigen retrieval was performed via heat mediation in a citrate buffer. Samples were blocked before incubating with primary antibody (1/100) for 1 hour at room temperature. An HRP-conjugated anti-mouse IgG was used as the secondary antibody and staining was detected using DAB.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labeling Mucin 5AC with ab3649 at 1/5000 dilution. Heat mediated antigen retrieval was performed using Citrate buffer pH 6 (epitope retrieval solution 1) for 20 minutes. The section was incubated with ab3649 for 30 mins at room temperature. Goat Anti-Mouse IgG H&L (HRP polymer) (Goat Anti-Mouse IgG H&L (HRP polymer) ab214879) was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
  

  The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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  Immunocytochemistry/ Immunofluorescence - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Immunocytochemistry analysis of A549 (human lung carcinoma cell line) cells labeling Mucin 5AC with ab3649 at 1/100 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-beta Tubulin rabbit monoclonal antibody (Anti-beta Tubulin antibody [EPR16774] ab179513) at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution (red). Nuclear DNA was labelled with DAPI (blue).
  

  Negative control 1: ab3649 (Mouse monoclonal antibody to Mucin 5AC at 1/100 dilution) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 (anti-Rabbit secondary Alexa Fluor^® 594 at 1/1000 dilution)

  Negative control 2: Anti-beta Tubulin antibody [EPR16774] ab179513 (Rabbit monoclonal antibody to beta Tubulin at 1/200 dilution) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 (anti-Mouse secondary Alexa Fluor^® 488 at 1/1000 dilution)

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  Immunohistochemistry (Frozen sections) - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Immunohistochemistry (Frozen sections) analysis of mouse stomach tissue labeling Mucin 5AC with ab3649 at 1/100 dilution. Tissue was fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Heat mediated antigen retrieval was performed using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) was used as the secondary antibody at 1/1000 dilution (green). Nuclei counterstained with DAPI (blue).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse stomach tissue labeling Mucin 5AC with ab3649 at 1/5000 dilution. Heat mediated antigen retrieval was performed using Citrate buffer pH 6 (epitope retrieval solution 1) for 20 minutes. The section was incubated with ab3649 for 30 mins at room temperature. Goat Anti-Mouse IgG H&L (HRP polymer) (Goat Anti-Mouse IgG H&L (HRP polymer) ab214879) was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
  

  The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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  Immunohistochemistry (Frozen sections) - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Immunohistochemistry (Frozen sections) analysis of rat stomach tissue labeling Mucin 5AC with ab3649 at 1/100 dilution. Tissue was fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Heat mediated antigen retrieval was performed using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) was used as the secondary antibody at 1/1000 dilution (green). Nuclei counterstained with DAPI (blue).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue labeling Mucin 5AC with ab3649 at 1/5000 dilution. Heat mediated antigen retrieval was performed using Citrate buffer pH 6 (epitope retrieval solution 1) for 20 minutes. The section was incubated with ab3649 for 30 mins at room temperature. Goat Anti-Mouse IgG H&L (HRP polymer) (Goat Anti-Mouse IgG H&L (HRP polymer) ab214879) was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
  

  The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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  Multiplex immunohistochemistry - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Fluorescence multiplex immunohistochemical analysis of the human stomach (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-Mucin 5AC (ab3649, gray; Opal™690), anti-MUC-6 (Anti-Gastric Mucin/MUC-6 antibody [EPR20623] ab223846, green; Opal™520) and anti-Rab3D (Anti-Rab3D antibody [EPR8106] ab128997, red; Opal™570) on human stomach. Panel B: anti-MUC-6 stained on mucous neck cells. Panel C: anti-Mucin 5AC stained on surface mucous cells. Panel D: anti-Rab3D stained on Chief cells. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab3649 (1/5000 dilution), Anti-Gastric Mucin/MUC-6 antibody [EPR20623] ab223846 (1/1000 dilution), and Anti-Rab3D antibody [EPR8106] ab128997 (1/10000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

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  This image is courtesy of an anonymous customer review

  Immunocytochemistry - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Immunocytochemistry analysis of paraformaldehyde-fixed 0.5% Triton in PBS-permeabilized human HBE cells staining with ab3649 at 1/300 dilution. Secondary antibody was Alexa Fluor® 594 Goat Thermo A-11005 at 1/250 dilution. Red: Actin Magenta: MUC5AC Blue: DNA. Samples were incubated with the primary antibody with 1% BSA in PBS for 16 hours at 4°C.

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  Immunocytochemistry/ Immunofluorescence - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Calu-3 (human lung adenocarcinoma epithelial cell) labelling (A) MUC5B with Anti-MUC5B antibody [EPR27200-159] ab315330 at 1/500 (1.008 ug/ml) 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 (2 ug/ml) dilution (Green), or (B) MUC5AC with ab3649 at 1/100 (10.45 ug/ml) followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

  This antibody does not cross-react with human MUC5AC.
  
  (A) No staining of MUC5B in Calu-3 cell line.
  
  (B) Positive staining of MUC5AC (ab3649, green) in Calu-3 cell line.
  
  
  

  Images were taken with a confocal microscope (Leica-Microsystems, TCS SP8).
  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 (Magenta). 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 (2 ug/ml) dilution.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mucin 5AC antibody [45M1] (ab3649)

  Immunohistochemical analysis of paraffin-embedded (A, C) A-549 (human lung carcinoma epithelial cell) cell pellet (B, D) Calu-3 (human lung adenocarcinoma epithelial cell) cell pellets.

  A & B: whole cell pellet labelling of Mucin 5B with Anti-MUC5B antibody [EPR27200-159] ab315330 at 1/100 dilution (5.04 ug/ml). C & D: whole cell pellet labelling of Mucin 5A with ab3649 at 1/5000 dilution (0.211 ug/ml).

  This antibody recognizes human MUC5B but not human MUC5AC.
  
  Positive staining on (A) A-549 cell pellet. No staining on (B) Calu-3 cell pellet. The sections were incubated with Anti-MUC5B antibody [EPR27200-159] ab315330 or ab3649 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-Mucin 5AC antibody [45M1] (ab3649)

  Immunohistochemical analysis of formalin fixed paraffin embedded human stomach labelling Mucin 5AC with ab3649 at a concentration of 0.05µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

  Anti-Mucin 5AC antibody [45M1] ab3649 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

  Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).