Product details
Antibody type = Mouse monoclonal / IgG1
Clone = HMV333
Positive control = Liver: A strong CA 19-9 staining should be seen in intrahepatic bile ducts (Note that reactivity of control tissues must be previously tested to exclude Le a-b-).
Negative control = Liver: CA 19-9 staining should be absent in all hepatocytes.
Cellular localization =Intracellular
Reactivity = Human
Application = Immunohistochemistry
Dilution = 1:100 – 1:200
Intended Use = Research Use Only
Relevance of Antibody
CA 19-9 is a commonly used serum tumor marker.
Biology Behind
Carbohydrate antigen 19-9 (CA19-9), also termed sialyl-Lewis A, is a cell surface glycoprotein complex which plays a role in cell-to-cell recognition processes. It is a tetrasaccharide carbohydrate containing a transmembrane protein skeleton and extracellular oligosaccharide chains which are extensively glycosylated. CA 19-9 expression is only possible in cells expressing the Lewis gene product, 1,4-fucosyltransferase. As this is only found in patients with Le (a-b+) or Le (a+b-) blood groups there are about 6% of Caucasians and 22% of non-Caucasians who cannot produce CA 19-9. Because CA 19-9 is particularly often overexpressed in pancreatic cancers, serum measurement of CA 19-9 has earlier been recommended for early detection of this cancer. Because of lack of specificity, this approach has largely been abandoned but CA19-9 serum measurement is still used as a marker for disease monitoring. CA19-9 is also being evaluated as a therapeutic target.
Staining Pattern in Normal Tissues
Images describing the CA 19-9 staining pattern in normal tissues obtained by the antibody HMV333 are shown in our “Normal Tissue Gallery”.
Brain | Cerebrum | Negative. |
Cerebellum | Negative. | |
Endocrine Tissues | Thyroid | Negative. |
Parathyroid | Negative. | |
Adrenal gland | Negative. | |
Pituitary gland | Negative. | |
Respiratory system | Respiratory epithelium | In some (but not all samples): Moderate to strong apical membranous CA 19-9 staining of most superficial epithelial cells. In addition, strong membranous and cytoplasmic positivity of a large fraction of goblet cells. |
Lung | Negative. | |
Gastrointestinal Tract | Salivary glands | Weak to strong, predominantly cytoplasmic (but also membranous) CA 19-9 staining of mucinous glandular cells. Moderate membranous staining of few myoepithelial cells.
Weak to moderate membranous CA 19-9 staining of few excretory duct cells. Serous glandular cells are largely negative. |
Esophagus | Strong membranous CA 19-9 staining of a significant fraction of cells in superficial layers of the squamous epithelium (not in all samples). | |
Stomach | Strong membranous and cytoplasmic CA 19-9 staining of a subset of cells in gastric glands. | |
Duodenum | Apical membranous CA 19-9 staining of few epithelial cells. | |
Small intestine | Weak to moderate apical membranous CA 19-9 staining of some superficial epithelial cells. Strong, predominantly membranous CA 19-9 staining of a significant fraction of goblet cells, especially at the surface of the epithelium. In addition, few epithelial cells – predominantly in the crypts – exhibit a strong cytoplasmic and membranous CA 19-9 positivity. | |
Appendix | Moderate to strong apical membranous CA 19-9 staining of most superficial epithelial cells. In addition, strong membranous and cytoplasmic positivity of a large fraction of goblet cells (not in all samples). | |
Colon | Weak to moderate, apical membranous CA 19-9 staining of some superficial epithelial cells. In addition, moderate to strong, predominantly membranous positivity of a fraction of goblet cells. | |
Rectum | Moderate membranous CA 19-9 staining of a small fraction of crypt epithelial cells. | |
Liver | Strong membranous and cytoplasmic CA 19-9 staining of intrahepatic bile ducts. Some hepatocytes can show a membranous CA 19-9 positivity at the apical/luminal membrane. | |
Gallbladder | Weak to strong, predominantly apical, membranous and cytoplasmic CA 19-9 staining of epithelial cells (not in all samples). | |
Pancreas | Strong, membranous and cytoplasmic CA 19-9 staining of intercalated ducts. | |
Genitourinary | Kidney | Weak to moderate, apical membranous CA 19-9 staining of few tubular and collecting duct cells. |
Urothelium | Weak to moderate membranous CA 19-9 staining of a fraction of urothelial cells, especially in the superficial cell layer. | |
Male genital | Prostate | Focal membranous CA 19-9 staining of epithelial cells (not in all samples |
Seminal vesicles | Negative. | |
Testis | Negative. | |
Epididymis | Weak to moderate CA 19-9 staining can be seen at the apical membrane of epithelial cells in the cauda. | |
Female genital | Breast | Moderate to strong, predominantly membranous CA 19-9 staining of luminal cells. The staining predominates at the apical membrane. |
Uterus, myometrium | Negative. | |
Uterus, ectocervix | Weak to moderate cytoplasmic CA 19-9 staining of a fraction of cells in the suprabasal and superficial layers of the squamous epithelium. | |
Uterus endocervix | Weak to strong, cytoplasmic CA 19-9 staining of variable intensity in some endocervical epithelial cells (not in all samples). | |
Uterus, endometrium | Weak to moderate CA 19-9 staining of few epithelial cells, predominantly at luminal membranes (in some samples). | |
Fallopian Tube | Moderate to strong CA 19-9 staining at the apical membrane of a fraction of epithelial cells. | |
Ovary | Negative. | |
Placenta early | Negative. | |
Placenta mature | Negative. | |
Amnion | Strong membranous and cytoplasmic CA 19-9 staining of a significant fraction of cells. | |
Chorion | Negative. | |
Skin | Epidermis | Negative. |
Sebaceous glands | Negative. | |
Muscle/connective tissue | Heart muscle | Negative. |
Skeletal muscle | Negative. | |
Smooth muscle | Negative. | |
Vessel walls | Negative. | |
Fat | Negative. | |
Stroma | Negative. | |
Endothelium | Negative. | |
Bone marrow/ lymphoid tissue | Bone marrow | Negative. |
Lymph node | Negative. | |
Spleen | Negative. | |
Thymus | Strong membranous and cytoplasmic CA 19-9 staining of most cells of corpuscles of Hassall‘s. | |
Tonsil | Weak to moderate membranous CA 19-9 staining of a fraction of cells in suprabasal and basal cell layers of the squamous epithelium. This can be seen in both surface and crypt epithelium. | |
Remarks |
A strong CA 19-9 staining occurs in various epithelial cell types such as for example in many goblet cells of the intestine, gastric glandular cells, gallbladder epithelium, intrahepatic bile ducts, mucinous cells in salivary and bronchial glands, and in luminal breast epithelial cells.
It is of note that – in line with an estimated 10% of analyzed samples being derived from Le a-b- patients – a complete absence of CA 19-9 staining occasionally occurs in all tissues.
Positive control = Liver: A strong CA 19-9 staining should be seen in intrahepatic bile ducts (Note that reactivity of control tissues must be previously tested to exclude Le a-b-).
Negative control = Liver: CA 19-9 staining should be absent in all hepatocytes.
Staining Pattern in Relevant Tumor Types
CA 19-9 is most expressed in pancreatic, colorectal, gastric, and esophageal adenocarcinomas, mucinous ovarian cancer, cervical adenocarcinoma, cholangiocarcinoma, and urothelial neoplasms, but can also be seen in various other tumor entities.
Compatibility of Antibodies
No data available at the moment
Protocol Recommendations
IHC users have different preferences on how the stains should look like. Some prefer high staining intensity of the target stain and even accept some background. Others favor absolute specificity and lighter target stains. Factors that invariably lead to more intense staining include higher concentration of the antibody and visualization tools, longer incubation time, higher temperature during incubation, higher temperature and longer duration of the heat induced epitope retrieval (slide pretreatment). The impact of the pH during slide pretreatment has variable effects and depends on the antibody and the target protein.
All images and data shown here and in our image galleries are obtained by the manual protocol described below. Other protocols resulting in equivalent staining are described as well.
Manual protocol
Freshly cut sections should be used (less than 10 days between cutting and staining). Heat-induced antigen retrieval for 5 minutes in an autoclave at 121°C in pH 7,8 Target Retrieval Solution buffer. Apply HMV333 at a dilution of 1:200 at 37°C for 60 minutes. Visualization of bound antibody by the EnVision Kit (Dako, Agilent) according to the manufacturer’s directions.
Potential Research Applications
- The diagnostic, prognostic, and predictive role of CA 19-9 expression in tumor tissue and in preneoplastic disease needs to be further clarified.
- The role of CA 19-9 as a therapeutic target is under investigation.
Evidence for Antibody Specificity in IHC
There are two ways how the specificity of antibodies can be documented for immunohistochemistry on formalin fixed tissues. These are: 1. Comparison with a second independent method for target expression measurement across a large number of different tissue types (orthogonal strategy), and 2. Comparison with one or several independent antibodies for the same target and showing that all positive staining results are also seen with other antibodies for the same target (independent antibody strategy).
Orthogonal validation: In case of CA 19-9 a comparison with RNA expression cannot be performed because CA 19-9 represents an aberrant glycosylation and not a genuinely expressed gene product.
Comparison of antibodies: True presence of CA 19-9 in all cell types for which CA 19-9 staining was observed by HMV333 is corroborated by confirmation of all these CA 19-9 positive cell types by a second independent commercially available anti-CA 19-9 antibody (termed “validation antibody”).