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Product details

Synonyms = CK13; Cytokeratin-13; Keratin Type I Cytoskeletal 13; Keratin-13; KRT13; Type I Cytoskeletal 13; WSN2

Antibody type = Mouse monoclonal / IgG1

Clone = MSVA-613M

Positive control = Tonsil: All squamous epithelial cells of the surface squamous epithelium (except the basal layer) and a fraction of squamous cells of the crypts should show strong KRT13 staining. 

Negative control = Tonsil: All lymphocytes and blood vessels must not show any KRT13 staining.

Cellular localization = Cytoplasmic

Reactivity = Human

 

Application = Immunohistochemistry
Dilution = 1:100 – 1:200
Intended Use = Research Use Only

Relevance of Antibody

Cytokeratin 13 is expressed in non-keratinizing squamous epithelial cells.

Biology Behind

Cytokeratin 13 (CK13), also termed keratin 13 (KRT13) is a type I acidic high molecular weight keratin protein encoded by the KRT13 gene located at 17q21. It dimerizes with the type I keratin 4 and forms intermediate filaments that primarily shape the cytoskeleton of specific epithelial cells. In these cells, KRT13 is part of the cytoskeletal scaffold which contributes to the cell architecture and provides the cells with the ability to withstand mechanical stress. Mutations in CK10 have been linked to the autosomal dominant disorder “White Sponge Nevus”.

Staining Pattern in Normal Tissues

Cytokeratin 13 staining pattern in Normal Tissues with antibody MSVA-613M (images 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 Very few KRT13 positive basal cells can occur.
Lung Negative.
Gastrointestinal Tract Salivary glands Very few KRT13 positive individual cells or small groups of cells can occur in the epithelium. Focal staining of cells and groups of cells in excretory ducts of salivary glands
Esophagus Strong CK13 staining of all supra-basal cell layers of non-keratinizing squamous epithelium.
Stomach Negative.
Duodenum Negative.
Small intestine Negative.
Appendix Negative.
Colon Negative.
Rectum Negative.
Liver Negative.
Gallbladder Faint staining of few gall bladder epithelial cells in a case with inflammation.
Pancreas Negative.
Genitourinary Kidney Negative.
Urothelium Strong CK13 staining of all urothelial cells except umbrella cells.
Male genital Prostate Very few KRT13 positive basal cells can occur.
Seminal vesicles Very few KRT13 positive basal cells can occur.
Testis Negative.
Epididymis Negative.
Female genital Breast Negative.
Uterus, myometrium Negative.
Uterus, ectocervix Strong CK13 staining of all supra-basal cell layers of non-keratinizing squamous epithelium.
Uterus endocervix Negative.
Uterus, endometrium Very few KRT13 positive individual cells or small groups of cells can occur in the epithelium.
Fallopian Tube Negative.
Ovary Negative.
Placenta early Negative.
Placenta mature Negative.
Amnion Cytoplasmic staining in few amnion cells.
Chorion Negative.
Skin Epidermis CK13 staining is usually lacking in keratinizing epithelium, but scattered KRT13 positive cells or groups of cells can occur.
Sebaceous glands Scattered KRT13 positive cells or groups of cells can occur in sebaceous glands and hair follicles.
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 CK13 staining of corpuscles of Hassall’s and of other epithelial cells in the medulla, but not in the cortex.
Tonsil Strong CK13 staining of all supra-basal cell layers of non-keratinizing squamous epithelium.
Remarks

 

These findings are largely consistent with the RNA and protein data summarized in the Human Protein Atlas (Tissue expression Cytokeratin 13) 

 

Suggested positive tissue control: Tonsil: All squamous epithelial cells of the surface squamous epithelium (except the basal layer) and a fraction of squamous cells of the crypts should show strong KRT13 staining. 

Suggested negative tissue control: Tonsil: All lymphocytes and blood vessels must not show any KRT13 staining.

 

Normal tissue gallery

Staining Pattern in Relevant Tumor Types

KRT13 immunostaining almost exclusively occurs in squamous cell carcinomas of various sites of origin. At frequencies <10% KRT13 expression can also be seen in adenocarcinomas of the pancreas, stomach, ovary, lung, endometrium, breast, esophagus, and the prostate, as well as in nonseminomatous germ cell tumors.

The TCGA findings on Cytokeratin 13 RNA expression in different tumor categories have been summarized in the Human Protein Atlas.

 

Cancer tissue gallery

 

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. Accordingly, multiple different protocols can generate identical staining results.

 

All images and data shown here and in our image galleries were 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 pH7,8 Target Retrieval Solution buffer. Apply MSVA-613M at a dilution of 1:150 at 37°C for 60 minutes. Visualization of bound antibody by the EnVision Kit (Dako, Agilent) according to the manufacturer’s directions.

 

 

 

Agilent / Dako – Autostainer Link 48

Pretreatment in PT-Link for 30 minutes at 95°C (pH high); FLEX peroxidase blocking for 5 minutes (room temperature), MSVA-613M 1:150 for 20 minutes (room temperature), FLEX+ mouse/rabbit (LINKER) for 15 minutes (room temperature), horseradish peroxidase (HRP) for 20 minutes (room temperature), FLEX DAB+Sub-Chromo for 10 minutes (room temperature), FLEX hematoxylin for 5 minutes (room temperature).

These images reflect stainings by the protocol described above. It is of note that a comparable staining result can also be obtained by different protocols. In general, a longer pretreatment, a longer incubation time of the primary antibody, a higher antibody concentration, and a longer incubation time of FLEX+LINKER result in stronger staining, potentially at the cost of more background staining. Modifications of the protocol with a strengthening effect on staining intensity in combination with changes of other parameters that result in lower staining intensity can result in a comparable result as shown above.

 

 

Leica – BOND RX

Dewax at 72°C for 30 seconds; Pretreatment in Bond Epitope Retrieval Solution (ER2 – EDTA pH9) for 20 minutes at 100°C; Peroxidase blocking for 5 minutes (room temperature), MSVA-613M 1:150 for 15 minutes (room temperature), Post primary (rabbit anti mouse) for 8 minutes (room temperature), Polymer (goat anti rabbit) for 8 minutes (room temperature), mixed DAB refine for 10 minutes (room temperature), hematoxylin for 5 minutes (room temperature).

These images reflect stainings by the protocol described above. It is of note that a comparable staining result can also be obtained by different protocols. In general, a longer pretreatment, a longer incubation time of the primary antibody, a higher antibody concentration, a higher temperature during incubation, and a longer incubation time of Post primary and or the Polymer result in stronger staining, potentially at the cost of more background staining. Modifications of the protocol with a strengthening effect on staining intensity in combination with changes of other parameters that result in lower staining intensity can result in a comparable result as shown above.

 

 

Roche – Ventana Discovery ULTRA

Pretreatment for 64 minutes at 100°C (pH 8,4); CM peroxidase blocking for 12 minutes (room temperature), MSVA-613M 1:150 for 20 minutes at 36°C, secondary antibody (anti-rabbit HQ) for 12 minutes at 36°C, anti-HQ HRP for 12 minutes at room temperature, DAB at room temperature, hematoxylin II at room temperature for 8 minutes, bluing reagent at room temperature for 4 minutes.

These images depict staining results obtained by the protocol described above. It is of note, that the Ventana machines generally require higher antibody concentrations than other commonly used autostainers because the antibodies are automatically diluted during the procedure. Various other protocols can result in an identical result as shown above. A longer pretreatment, a longer incubation time of the primary antibody, a higher antibody concentration, a higher temperature during incubation, and a longer incubation time of secondary antibody and or the anti-HQ HRP result in stronger staining, potentially at the cost of more background staining.

 

Impact of pH

 

The strongest KRT13 staining by MSVA-613M is obtained at a pH 9,0. However, pH 7,8 results in only a slight reduction of the staining intensity as compared to pH9. We thus consider pH7,8 as optimal for manual staining because of the better tissue preservation at pH7,8 than at pH 9,0.

Potential Research Applications

  • The diagnostic utility of KRT713 expression analysis should be investigated in a large cohort of tumors from different entities.
  • The diagnostic utility of increased KRT13 expression in the skin is not known.
  • The diagnostic utility of decreased KRT13 expression in non-keratinizing squamous epithelium of ectocervix, esophagus, oral cavity and lip is not known.
  • The prognostic role of KRT13 expression in squamous cell carcinoma is unknown.

Evidence for Antibody Specificity in IHC

Utility of MSVA-613M is documented by a staining pattern that exactly matches the data summarized in the protein atlas. A strong positive KRT13 staining is almost exclusively seen in non-keratinizing squamous epithelial cells but not in the skin. In addition, all tissues known to not express KRT13 including those notorious for non-specific IHC background such as kidney and colonic mucosa are completely KRT13 negative.

 

Normal tissue gallery