Literature references :
Reis CA, Sorensen T, Mandel U, David L, Mirgorodskaya E, Roepstorff P, Kihlberg J, Hansen JE, Clausen H.  Development and characterization of an antibody directed to an alpha-N-acetyl-D-galactosamine glycosylated MUC2 peptide.
Glycoconj J 1998 Jan;15(1):51-62

Isotype : IgM

Origin, source : A synthetic peptide (PTTTPISTTTMVTPTPTPTC) based on the sequence of  the tandem repeat of Muc2 was terminally glycosylated with GalNAc and miice were immunised with the glycosylated Gal-NAc Muc2 peptide coupled with KLH. The parental cells used for the fusion were NS1.

Specificity : PMH1 recognises specifically the glycosylate GalNAc-MUC2 peptide. It shows a positive signal with the petide glycosylated with a minimun of 1 mol of GalNAc attached. The epitope of the PHM1 antibody is located at the Carboxy-terminal end of the peptide.

Cross-reactivity : The PMH1 monoclonal antibody does not react with the unglycosylated peptide and does not cross-react with the Tn antigen or a Gal-NAc MUC1 peptide.

Binding Assays : PMH1 antibody reacts with the native and  the deglycosylated (TFMSA and neuraminidase treatment) mucin extracted from LS174T.

Immunoblotting : PMH1 antibody reacts strongly with the deglycosylated (TFMSA and neuraminidase treatment) mucin extracted from LS174T, a weak reaction was observed with the native mucin.

Immunohistochemistry:
Experiments performed on frozen tissues:
 

Positive tissues

Negative tissues

Salivary gland (supranuclear staining of mucus acinar cells from labial salivary gland) Stomach body  (supranuclear staining of foveolar epithelium) Stomach antrum (supranuclear staining of foveolar epithelium, diffuse cytoplasmic staining of cells of the pyloric glands) Stomach-intestinal metaplasia (strong staining of vacuoles and mucus goblet cells and supranuclear staining of columnar cells  ) Gallbladder (supranuclear staining of epithelial cells) Pancreas (supranuclear staining of acinar cells and duct cells ) Small intestine (strong supranuclear staining of goblet cells) Colon (strong staining of vacuoles and mucus in goblet cells) Bronchus (supranuclear staining of bronchial cells) Kidney (apical membrane staining of tubular cells)

Breast Liver Ovary Spleen Uterus

Experiments performed in Jacques Bara's lab:

The PMH1 antibody did not work on ethanol fixed tissues.
 

Other publications using the antibody :
Reis CA, David L, Correa P, Carneiro F, de Bolos C, Garcia E, Mandel U, Clausen H, Sobrinho-SimoesM.  Intestinal metaplasia of human stomach displays distinct patterns of mucin (MUC1, MUC2, MUC5AC, and MUC6) expression.
Cancer Res. 1999 Mar 1;59(5):1003-7.

Carvalho F, David L, Aubert JP, Lopez-Ferrer A, De Bolos C, Reis CA, Gartner F, Peixoto A, Alves P, Sobrinho-Simoes M.  Mucins and mucin-associated carbohydrate antigens expression in gastric carcinoma cell lines.
Virchows Arch 1999 Nov;435(5):479-85

 Machado JC, Nogueira AM, Carneiro F, Reis CA, Sobrinho-Simoes M.  Gastric carcinoma exhibits distinct types of cell differentiation: an immunohistochemical study of trefoil   peptides (TFF1 and TFF2) and mucins (MUC1, MUC2, MUC5AC, and MUC6).
J Pathol. 2000 Mar;190(4):437-43.

Nogueira AM, Machado JC, Carneiro F, Reis CA, Gott P, Sobrinho-Simoes M.  Patterns of expression of trefoil peptides and mucins in gastric polyps with and without malignant  transformation.
J Pathol. 1999 Apr;187(5):541-8.

Note : Made in H.Clausen Laboratory