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Product details

Synonyms = Cadherin-16 (CDH16); Kidney-specific cadherin; Ksp-cadherin antibody

Antibody type = Recombinant Rabbit monoclonal / Rabbit IgG

Clone = MSVA-516R

Positive control =  Kidney: A strong staining should be seen in distal tubuli and collecting ducts while the staining is at least moderate in proximal tubuli. 

Negative control = Colon: All cells should be CDH16 negative. 

Cellular localization = Cell Surface and Cytoplasmic

Reactivity = Human

 

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

Relevance of Antibody

CDH16 is a marker for kidney and thyroid epithelial cells.

Biology Behind

Cadherin-16, coded by the CDH16 gene at 16q22.1, belongs to the cadherin superfamily which is composed of calcium-dependent, membrane-associated glycoproteins with a role in cell-adhesion. CDH16 is involved in embryonal development and cell growth. CDH16 supports the formation of tubuli during renal development and remains expressed in distal tubuli of adult kidneys. CDH16 is also relevant for the development of follicular thyroid cells and thyroid follicular polarity.

Staining Pattern in Normal Tissues

CDH16 staining pattern in Normal Tissues with antibody MSVA-516R (images are shown in our “Normal Tissue Gallery”)

Brain Cerebrum Negative.
Cerebellum Negative.
Endocrine Tissues Thyroid Strong predominantly membranous CDH16 staining of follicular cells.
Parathyroid Negative.
Adrenal gland Negative.
Pituitary gland Negative.
Respiratory system Respiratory epithelium Negative.
Lung Negative.
Gastrointestinal Tract Salivary glands Negative.
Esophagus Negative.
Stomach Negative.
Duodenum Negative.
Small intestine Negative.
Appendix Negative.
Colon Negative.
Rectum Negative.
Liver Negative.
Gallbladder Moderate to strong, predominantly membranous CDH16 staining of a small fraction of epithelial cells which are often arranged in groups.
Pancreas Negative.
Genitourinary Kidney Strong predominantly membranous but also cytoplasmic CDH16 staining of distal tubuli and collecting ducts. Staining is somewhat weaker and predominantly seen in basal and lateral membranes of proximal tubuli.
Urothelium Negative.
Male genital Prostate Negative.
Seminal vesicles Moderate to strong, predominantly membranous CDH16 staining of a small fraction of epithelial cells which are often arranged in groups.
Testis Negative.
Epididymis Strong, predominantly membranous but also cytoplasmic CDH16 staining in all epithelial cells of the cauda epididymis. CDH16 is very weak or absent in epithelial cells of the caput.
Female genital Breast Negative.
Uterus, myometrium Negative.
Uterus, ectocervix Negative.
Uterus endocervix Moderate to strong, predominantly membranous CDH16 staining of a small fraction of epithelial cells which are often arranged in groups.
Uterus, endometrium Moderate to strong, predominantly membranous CDH16 staining of a small fraction of epithelial cells which are often arranged in groups.
Fallopian Tube Moderate to strong, predominantly membranous CDH16 staining of a small fraction of epithelial cells which are often arranged in groups.
Ovary Negative.
Placenta early Negative.
Placenta mature Negative.
Amnion Negative.
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 Negative.
Tonsil Negative.
Remarks

The findings described above are this consistent with the RNA data described in the Human Protein Atlas (Tissue expression CDH16)

 

Positive control =  Kidney: A strong staining should be seen in distal tubuli and collecting ducts while the staining is at least moderate in proximal tubuli. 

Negative control = Colon: All cells should be CDH16 negative. 

 

Normal tissue gallery

Staining Pattern in Relevant Tumor Types

A positive CDH16 immunostaining is most commonly seen in kidney cancer. CDH16 expression also occurs in cancers of the thyroid, uterine cervix, endometrium and the ovary.

The TCGA findings on CDH16 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.

 

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 MSVA-516R 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-516R 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-516R 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.

 

Potential Research Applications

  • The diagnostic utility of CDH16 immunostaining should be evaluated in studies comparing CDH16 staining in a broad range of different cancers.
  • The prognostic role of CDH16 expression loss should be investigated in cancers of the kidney and of the thyroid.