Isotype : IgG1
Literature references :
Reis CA, David L, Nielsen PA, Clausen H, Mirgorodskaya K, Roepstorff
P, Sobrinho-Simoes M. Immunohistochemical study of MUC5AC expression
in human gastric carcinomas using a novel monoclonal antibody.
Int J Cancer 1997 Feb 20;74(1):112-21
Origin, source : Mice were immunised with a MUC5AC tandem repeat peptide (CTTSTTSAPTTSTTSAPTTS). The sera were tested dy ELISA, immunihistology and Western blot. NS-1 cells were used as parental cells for the fusion.
Specificity : The CLH2 antibody reacts with three tandem repeat peptides (TTSTTSAP, SAPTTSTTSAPT, CTTSTTSAPTTSTTSAPTTS). Western blot with the native and deglycosylated mucin extract from gastric carcinoma were used to screen the hybridomas.
Cross-reactivity : No cross reactivity with other peptides from MUC2, MUC3, MUC6 and MUC7.
Binding Assays : Direct, indirect and inhibitory ELISA techniques were used for the screening of the hybridomas using the MUC5AC peptides and other irrevelant peptides. CLH2 monoclonal antibody shows no difference of reactivity with the three MUC5AC peptides. The antibody reacts also with a high molecular weight fraction from gel centrifugation of mucin extract from normal gastric antrum and body as well as gastric carcinoma.
Immunoblotting : The CLH2 antibody shows a weak reactivity with a high molecular weight smear characteristic of mucin from native mucin extract from normal gastric mucosa and from gastric diffuse carcinoma .
Results of immunohistochemistry and experiments performed in Jacques Bara's lab
:
Tissues | Immunohistology | ISH | |
Oesophageal glands | - | ND | |
Cardia | Surface
Glands |
+++
- |
ND
ND |
Fundus | Surface
Glands |
+++
- |
+++
- |
Pylorus | Surface
Glands |
+++
- |
+++
- |
Duodenum | Villi
Brunner's glands |
-
- |
-
- |
Jejunum | Villi
Glands |
-
- |
-
- |
Ileum | Villi
Glands |
-
- |
-
- |
Colon | - | - | |
Prostate | utriculus | + | ??? |
Conjunctiva | - | ++++ | |
Endocervix | - | ++ | |
Respiratory tract | Surface
Glands mucus cells Serous cells |
++++
- - |
+++
+++/-* - |
Bile duct | - | - | |
Pancreas | Islets of Langherans | - | - |
Pancreas | Wirsung duct
Small ducts |
-
- |
ND
- |
Gall Bladder | +/- | + |
* Carme De Bolos results
Results of immunohistochemistry experiments
from the original publication :
Samples fixed in formalin and embedded in paraffin.
Normal tissues
CLH2 positive | CLH2 negative |
Bronchus (Ciliated epithelium,
supranuclear cytoplasm)
Endocervix (Superficial epithelium-few cells, supranuclear cytoplasm) Stomach body (Foveolar epithelium-perinuclear and diffuse cytoplasmic, mucous neck cells-perinuclear) Stomach antrum (Foveolar epithelium-perinuclear and diffuse, mucous neck cells-perinuclear) Gall bladder |
Salivary glands
Esophagus Thyroid Stomach body (Oxynthic glands) Stomach antrum (Ryloric glands) Breast Pancreas Liver Small intestine Colon Exocervix Kidney Ovary Prostate Skin Spleen |
Other publications using the antibody:
Van den Brink GR, Tytgat KM, Van der Hulst RW, Van der Loos CM,
Einerhand AW, Buller HA, Dekker J. H pylori colocalises with
MUC5AC in the human stomach.
Gut. 2000 May;46(5):601-7.
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.
Notes : Made in the Clausen Laboratory