IMMUNOHISTOCHEMISTRY CATHEPSIN D Test
Test Name: IMMUNOHISTOCHEMISTRY CATHEPSIN D Test
Components: Cathepsin D Test
Price: 410.0 AED
Sample Condition: Submit tumor tissue in 10% Formal-saline OR Formalin fixed paraffin embedded block. Ship at room temperature. Provide a copy of the Histopathology report, Site of biopsy and Clinical history.
Report Delivery: Sample Daily by 6 pm; Report Block: 5 days Tissue Biopsy: 5 days Tissue large complex: 7 days
Method: Immunohistochemistry
Test type: Cancer
Doctor: Oncologist, Pathologist
Test Department: DNA Labs UAE
Pre Test Information: Provide a copy of the Histopathology report, Site of biopsy and Clinical history.
Test Details
Immunohistochemistry (IHC) is a technique used to visualize specific proteins or antigens in tissue samples. Cathepsin D is a lysosomal protease that plays a role in protein degradation and is often used as a marker for lysosomal activity.
To perform an immunohistochemistry Cathepsin D test, the following steps are typically followed:
- Tissue preparation: Tissue samples, usually obtained from biopsies or surgical resections, are fixed in formalin and embedded in paraffin wax. The tissue is then cut into thin sections (usually around 4-6 micrometers thick) using a microtome.
- Deparaffinization and rehydration: The paraffin wax is removed from the tissue sections by incubating them in xylene or other clearing agents. The sections are then rehydrated by passing them through a series of graded alcohols.
- Antigen retrieval: Formalin fixation can cause cross-linking of proteins, making them inaccessible to antibodies. Antigen retrieval methods, such as heat-induced epitope retrieval or enzymatic digestion, are used to unmask the antigenic sites and improve antibody binding.
- Blocking: To prevent non-specific binding of antibodies, the tissue sections are incubated in a blocking solution containing proteins (e.g., bovine serum albumin or non-fat milk) or other blocking agents.
- Primary antibody incubation: The tissue sections are incubated with a primary antibody specific to Cathepsin D. The primary antibody binds to the target protein in the tissue sections.
- Secondary antibody incubation: After washing off unbound primary antibody, the tissue sections are incubated with a secondary antibody conjugated to a detectable label (e.g., enzyme, fluorophore, or chromogen). The secondary antibody binds to the primary antibody, amplifying the signal.
- Visualization: The detectable label attached to the secondary antibody generates a visible signal, allowing the visualization of Cathepsin D in the tissue sections. The signal can be visualized using various techniques, such as enzyme-substrate reactions (e.g., DAB) or fluorescent microscopy.
- Counterstaining and mounting: To enhance tissue structure visualization, counterstaining with dyes like hematoxylin may be performed. Finally, the tissue sections are mounted on glass slides with a mounting medium.
- Interpretation: The stained tissue sections are examined under a microscope by a pathologist or researcher. The presence and distribution of Cathepsin D can be assessed, and the staining intensity or pattern can provide insights into its role in the tissue. It’s important to note that specific protocols and reagents may vary depending on the laboratory and the specific Cathepsin D antibody used. Additionally, appropriate positive and negative controls should be included in the test to ensure accurate interpretation of the results.
Test Name | IMMUNOHISTOCHEMISTRY CATHEPSIN D Test |
---|---|
Components | |
Price | 410.0 AED |
Sample Condition | Submit tumor tissue in 10% Formal-saline OR Formalin fixed paraffin embedded block. Ship at room temperature. Provide a copy of the Histopathology report, Site of biopsy and Clinical history. |
Report Delivery | Sample Daily by 6 pm; Report Block : 5 days Tissue Biopsy : 5 days Tissue large complex : 7 days |
Method | Immunohistochemistry |
Test type | Cancer |
Doctor | Oncologist, Pathologist |
Test Department: | |
Pre Test Information | Provide a copy of the Histopathology report, Site of biopsy and Clinical history. |
Test Details | Immunohistochemistry (IHC) is a technique used to visualize specific proteins or antigens in tissue samples. Cathepsin D is a lysosomal protease that plays a role in protein degradation and is often used as a marker for lysosomal activity. To perform an immunohistochemistry Cathepsin D test, the following steps are typically followed: 1. Tissue preparation: Tissue samples, usually obtained from biopsies or surgical resections, are fixed in formalin and embedded in paraffin wax. The tissue is then cut into thin sections (usually around 4-6 micrometers thick) using a microtome. 2. Deparaffinization and rehydration: The paraffin wax is removed from the tissue sections by incubating them in xylene or other clearing agents. The sections are then rehydrated by passing them through a series of graded alcohols. 3. Antigen retrieval: Formalin fixation can cause cross-linking of proteins, making them inaccessible to antibodies. Antigen retrieval methods, such as heat-induced epitope retrieval or enzymatic digestion, are used to unmask the antigenic sites and improve antibody binding. 4. Blocking: To prevent non-specific binding of antibodies, the tissue sections are incubated in a blocking solution containing proteins (e.g., bovine serum albumin or non-fat milk) or other blocking agents. 5. Primary antibody incubation: The tissue sections are incubated with a primary antibody specific to Cathepsin D. The primary antibody binds to the target protein in the tissue sections. 6. Secondary antibody incubation: After washing off unbound primary antibody, the tissue sections are incubated with a secondary antibody conjugated to a detectable label (e.g., enzyme, fluorophore, or chromogen). The secondary antibody binds to the primary antibody, amplifying the signal. 7. Visualization: The detectable label attached to the secondary antibody generates a visible signal, allowing the visualization of Cathepsin D in the tissue sections. The signal can be visualized using various techniques, such as enzyme-substrate reactions (e.g., DAB) or fluorescent microscopy. 8. Counterstaining and mounting: To enhance tissue structure visualization, counterstaining with dyes like hematoxylin may be performed. Finally, the tissue sections are mounted on glass slides with a mounting medium. 9. Interpretation: The stained tissue sections are examined under a microscope by a pathologist or researcher. The presence and distribution of Cathepsin D can be assessed, and the staining intensity or pattern can provide insights into its role in the tissue. It’s important to note that specific protocols and reagents may vary depending on the laboratory and the specific Cathepsin D antibody used. Additionally, appropriate positive and negative controls should be included in the test to ensure accurate interpretation of the results. |