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cancer biomarkers guide

Cancer biomarker guide

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              • IHC guide
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                      • Biomarker antibody validation

                        Use our cancer immunohistochemistry (IHC) guide to biomarkers to select the right biomarker and the best IHC antibody for your research.



                        Overview

                        • The role of biomarkers in cancer research
                        • Immunohistochemistry in cancer biomarker research
                        • Choosing an antibody for IHC
                        • Find your guide
                          ​

                        ​The role of biomarkers in cancer research

                        Biomarkers play a pivotal role in cancer research. Whether this is identifying new biomarkers for improved diagnostics and drug development, validating current biomarkers across different cancer types and stages, or prioritizing targets for translation to preclinical research, the impact of cancer biomarkers on the progression and implementation of the research of these diseases has been vast. Continuing research in this field offers hope for new biomarkers and increased validation and applications of current biomarkers to improve diagnostics and therapeutics for all cancer types. View our video below to find out more. 

                        Download a pdf version of this guide here.

                          Estrogen receptor alpha
                          Determining the distribution of estrogen receptor alpha (ER-α) in breast cancer samples is an important initial step for the diagnosis and treatment evaluation of the disease (Hicks et al., 2017). Approximately 70% of breast cancer samples will give a positive staining signal for ER-α making it a crucial biomarker for breast cancer diagnosis (Jeselsohn et al., 2015). ER-α is a nuclear protein with a ligand-dependent transcription factor function. It is also most commonly detected in both luminal A and B subtypes of breast cancer (Jeselsohn et al., 2015).
                          IHC stain: nuclear signal
                          Recommended IHC antibody: Recombinant Anti-Estrogen Receptor alpha antibody [EPR4097] - ChIP Grade (ab108398)
                          Progesterone receptor
                          The progesterone receptor (PR) is another biomarker important for the initial diagnosis and evaluation of breast cancer (Hicks et al., 2017). PR is known to be induced by ER-α and plays a key role in ER-α protein regulation. PGR as a biomarker is therefore commonly used as an indicator of ER-α function (Mohammed et al., 2015). PR is highly expressed in luminal A-type breast cancer tissue and is associated with a good prognosis (Lim et al., 2016). 
                          IHC stain: nuclear signal
                          Recommended IHC antibody: Recombinant Anti-Progesterone Receptor antibody [YR85] (ab32085)
                          ErbB2/HER2
                          Overexpression of ErbB2/HER2 can be found in 20–30% of breast cancer tumors and is commonly found in more aggressive types of the disease (Mitri et al., 2012). ErbB2/HER2 is used as a diagnostic indicator for the FDA approved monoclonal antibody therapy, Trastuzumab (Herceptin). During this therapy ErbB2/HER2 acts as the binding target of Trastuzumab (Piccart-Gebhart et al., 2006). 
                          IHC stain: cytoplasmic, nuclear, and strongly in the cell membrane
                          Recommended IHC antibody: Recombinant Anti-ErbB 2 antibody [SP3] (ab16662)
                          p53
                          Mutated versions of the tumor suppressor p53 can be found in 80% of triple-negative breast cancer (TNBC) cases, ie tumors lacking expression of ER, PR, and ErbB2/HER2 (Duffy et al., 2018). This makes mutant p53 an excellent biomarker for TNBC, one of the more difficult breast cancers to treat as it will not respond to endocrine or anti-HER2 treatments (Li et al., 2019). Currently, there are mixed findings on the prognostic potential of p53 expression in breast cancer. Some studies have associated it with a positive prognostic outcome and others negative. (Li et al., 2019). p53 IHC stains are commonly used for diagnostic purposes.
                          IHC stain: cytoplasm and strong nuclear
                          Recommended IHC antibody: Anti-p53 antibody [DO-1] - ChIP Grade (ab1101)
                          BRCA1
                          Mutations in the BRCA genes are some of the most well-known breast cancer-associated protein mutations. BRCA1 is a tumor suppressor gene and one of the most commonly mutated genes in breast cancer. Patients with a BRCA1 mutation have a 40–80% increased risk of developing the disease making it a crucial biomarker in the diagnosis and prognosis of many breast cancer types (Fackenthal et al., 2007). 
                          IHC stain: nuclear signal
                          Recommended IHC antibody: Anti-BRCA1 antibody [MS110] (ab16780)
                          Epidermal growth factor receptor (EGFR)
                          EGFR is a transmembrane receptor and its expression is frequently found in TNBC and inflammatory breast cancer (IBC). It is most commonly associated with a poor prognosis (Masuda et al., 2012). High levels of EGFR expression is linked to an increased ability of the breast cancer to undergo metastasis making it a common biomarker for aggressive breast cancer types (Ali et al., 2016).  
                          IHC stain: cytoplasm and strong nuclear
                          Recommended IHC antibody: Anti-EGFR antibody [EP38Y] (ab52894)
                          CK20 (Cytokeratin 20) and CK7 (Cytokeratin 7)
                          CK7 and CK20 are cytokeratins expressed in the breast epithelia. Different CK7 and CK20 IHC expression patterns are commonly used to distinguish many carcinoma types including breast carcinomas. Most breast cancers are CK7 positive and CK20 negative, making the combination of these cytokeratins an excellent biomarker combination. Approximately 80% of breast adenocarcinomas are CK7 positive and CK20 negative (Chu et al., 2000). 
                          IHC stain: cytoplasm 
                          Recommended CK20 IHC antibody: Recombinant Anti-Cytokeratin 20 antibody [EPR1622Y] - Cytoskeleton Marker (ab76126) 
                          Recommended CK7 IHC antibody: Recombinant Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (ab68459)
                          APOBEC3B (apolipoprotein B mRNA editing catalytic polypeptide-like 3B)
                          APOBEC3B is expressed in ER-positive breast cancers and is used as a biomarker of poor prognosis for the disease and is a driver for downstream genetic mutations which accelerate the progression of breast cancer. These mutations make APOBEC3B positive breast cancers extremely drug-resistant (Zou et al., 2017). 
                          IHC stain: nuclear signal
                          Recommended IHC antibody: Anti-APOBEC3B antibody - N-terminal (ab191695)
                          Cell type-specific markers
                          FOXA1
                          The transcription factor FOXA1 has been shown to have a unique distribution within breast cancer cells compared to other cell types. FOXA1 binding is crucial for chromatin opening and the transcriptional activation of ER-α responsive genes within breast cancer cells (Cowper-Sallari et al., 2012). Studies have identified thousands of breast cancer-associated single nucleotide polymorphisms (SNPs) within the enhancer regions for FOXA1 (Meyer et al., 2012). 
                          IHC stain: nuclear signal
                          Recommended IHC antibody: Recombinant Anti-FOXA1 antibody [EPR10881] (ab170933)
                          GATA-binding protein 3 (GATA3)
                          GATA3 is a zinc finger transcription factor crucial for breast luminal epithelium differentiation. It is also a diagnostic biomarker for both primary and metastatic breast cancer and commonly found in luminal A and B breast cancer tissue. GATA3 also has a strong association with HER2 positive and ER- α positive hormone response in luminal breast cancers (Shaoxian et al., 2017). 
                          IHC stain: nuclear signal
                          Recommended IHC antibody: Recombinant Anti-GATA3 antibody [EPR16651] - ChIP Grade (ab199428)
                          CK5 (Cytokeratin 5)
                          CK5 is a cytokine found to be expressed in the basal cell layer of the mammary duct. Tumors that arise from these cells are also known to be CK5 positive and so CK5 is an excellent biomarker for the diagnosis of basal type breast cancers. The CK5 positive progenitor cells may also differentiate into glandular and myoepithelial cancer types (Laakso et al., 2005).  
                          IHC stain: cytoplasm and cell membrane
                          Recommended IHC antibody: Recombinant Anti-Cytokeratin 5 antibody [EP1601Y] - Cytoskeleton Marker (ab52635)
                          Smooth muscle myosin heavy chain (SMMHC)
                          SMMHC is expressed specifically in the contractile myoepithelial cells of the breast. A loss of the myoepithelial layer is commonly associated with invasive breast cancers. Markers such as SMMHC can be used in IHC to detect this layer more reliably than a hematoxylin and eosin stain alone (Zaha et al., 2014). 
                          IHC stain: cytoplasm
                          Recommended IHC antibody: Recombinant Anti-smooth muscle Myosin heavy chain 11 antibody [EPR5336(B)] (ab133567)
                          E-cadherin
                          E-cadherin is expressed at the cell junction of normal breast epithelial cells. A loss of E-cadherin is associated with a phenotypic switch by these cells to become invasive and migratory breast cancer cells. Loss of E-cadherin expression is commonly used as a biomarker for metastatic lobular breast carcinomas (Singhai et al., 2011).
                          IHC stain: Cell junctions
                          Recommended IHC antibody: Recombinant Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)
                          Alpha smooth muscle actin (ACTA2)
                          Smooth muscle actin is highly expressed in normal breast myoepithelial cells. It is commonly used as a marker of this normal breast cell type. Smooth muscle actin is commonly used for histology in conjunction with SMMHC and calponins to mark out the myoepithelial layer. A combination of markers such as this is much more reliable than using hematoxylin and eosin stains alone (Zaha et al., 2014).
                          IHC stain: cytoplasm
                          Recommended IHC antibody: Anti-alpha smooth muscle Actin antibody (ab5694)
                          Calponin 
                          Calponins are proteins found within the contractile components of the myoepithelium. Similarly to SMMHC and ACTA2, calponin stains are used as a marker of the myoepithelial layer in the diagnosis of invasive breast cancers (Zaha et al., 2014). 
                          IHC stain: cytoplasm
                          Recommended IHC antibody: Recombinant Anti-Calponin 1 antibody [EP798Y] (ab46794), Anti-Calponin 2 antibody (ab129331), Anti-Calponin 3 antibody (ab151427)
                          Delta-1-Catenin 
                          Delta-1-Catenin is a member of the p120 family and acts as a binder of E-cadherin. Delta-1-Catenin can be used in histology to detect invasive lobular breast cancer, high tumor-node-metastasis stage, and lymph node metastasis. It is also strongly associated with HER2 positive breast cancers and can be used as an indicator of poor prognosis as it is thought to promote a malignant phenotype (Zhang et al., 2015). 
                          IHC stain: cytoplasm and cell membrane
                          Recommended IHC antibody: Recombinant Anti-delta 1 Catenin/CAS antibody [EPR357(2)] (ab92514)
                          CK14 (Cytokeratin 14)
                          CK14 is a cytokine found to be expressed in the basal cell layer of the mammary duct. It is most commonly found to be co-expressing with CK5 is this tissue type and similarly to CK5, the histological detection of CK14 is an excellent biomarker for the diagnosis of basal type breast cancers (Laakso et al., 2005).
                          IHC stain: cytoplasm
                          Recommended IHC antibody: Recombinant Anti-Cytokeratin 14 antibody [EPR17350] - Cytoskeleton Marker (ab181595)
                          Other markers
                          Ki67
                          Ki67 is an important protein involved in cell division and is commonly used as a marker of cellular proliferation. It is also used in breast cancer diagnosis to determine the level of cell proliferation which can be a prognostic marker and a good indication of well certain breast cancers will respond to endocrine therapy (Niazi et al., 2018). 
                          IHC stain: nuclear
                          Recommended IHC antibody: Anti-Ki67 antibody (ab15580)
                          Cyclin D1
                          Cyclin D1 is one of the main regulatory proteins of the cell cycle promoting the cell cycle progression from G1 to S phase. Mutations in cyclin D1 are amplified in many different types of cancer including many breast cancers. 50% of breast cancers will contain an overexpression of cyclin D1 (Mohammadizadeh et al., 2013). It has been used as a biomarker for poor prognosis for many breast cancer types, but some studies are beginning to suggest that it could also suggest a positive outcome if expressed in some luminal subtypes of breast cancer (Ortiz et al., 2017). 
                          IHC stain: nuclear
                          Recommended IHC antibody: Recombinant Anti-Cyclin D1 antibody [EPR2241] - C-terminal (ab134175)
                          Cyclooxygenase-2 (COX-2)
                          COX-2 is an enzyme responsible for producing prostanoids. Its expression in breast tissue correlates strongly with breast cancer development. Overexpression of COX-2 has been shown to drive breast cancer phenotypes and blocking the action of this protein also shows potential for breast cancer therapy. There is also a strong link between COX-2 overexpression in breast adipose tissue and cancer progression linked to obesity (Harris et al., 2014). 
                          IHC stain: cytoplasm
                          Recommended IHC antibody: Recombinant Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
                          Caveolin-1
                          Caveolin-1 is the main component of caveolae, small invaginations found in the cell membrane. It’s also known to play a key role in cell proliferation, invasion, and breast cancer metastasis. It is also used as a biomarker for breast cancer treatments and disease outcome. High expression of caveolin-1 is a sign of poor prognosis an indicates are more aggressive metastatic breast cancer (Qian et al., 2019). 
                          IHC stain: cell membrane
                          Recommended IHC antibody: Recombinant Anti-Caveolin-1 antibody [E249] - Caveolae Marker (ab32577)
                          Peroxisome proliferator-activated receptor (PPAR-γ)
                          PPAR-γ is a well-studied nuclear hormone receptor. PPAR-γ has been implicated in different types of breast cancer in which its ligands inhibit proliferation and angiogenesis and induce apoptosis (Fenner et al., 2005). It has also been shown to play a role in promoting breast tumor growth through the maintenance of HER2 positive endothelial breast cancer stem cells making its expression a good biomarker for this cell type (Wang et al., 2013). 
                          IHC stain: nuclear
                          Recommended IHC antibody: Anti-PPAR gamma antibody - ChIP Grade (ab45036)

                          ​​

                          Immunohistochemistry in cancer biomarker research

                          IHC is a hugely popular tool in cancer research and remains the gold standard in clinical cancer diagnostics. Each different type of cancer is associated with specific protein biomarkers that can be identified via IHC to offer a powerful tool to investigate tumor development and responses. Using this technique researchers can identify the histogenesis of tumors, stage tumors, predict tumor prognosis, and predict and monitor therapeutic response. Knowing the utilities and pitfalls of each tumor-associated biomarker is essential to avoid potential diagnostic error, as no absolutely cancer-specific biomarker exists.

                          These tests can be run quickly, with ease, and at low cost compared to other diagnostic methods. IHC is thus a robust and reliable technique and, with a good panel of specific biomarkers, it is ideal for rapid clinical diagnosis.

                          Choosing an antibody for IHC

                          Whatever cancer type you are interested in, an essential starting point is the use of a sensitive, specific and consistent IHC antibody. Antibody validation is key to ensuring antibody specificity and reproducibility. We use a variety of experimental applications to validate our antibodies, including IHC and many of the antibodies on our catalog will state that they are suitable for use in IHC.

                          For consistent results between your IHC experiments and to ensure the long-term viability of your research we recommend using an antibody generated using recombinant technology (i.e. recombinant antibodies). These antibodies are developed in vitro and do not rely on an animal's immune system for production, meaning that batch-to-batch they deliver consistency of performance.

                          In this guide, we have included a number of the most common cancer types: breast, lung, colorectal, prostate, ovarian and pancreatic cancer and a range of emerging and approved biomarkers for diagnosis, prognosis and therapeutic monitoring.

                          Here you will be able to find the right biomarker for your research and select the most highly recommended antibody for use in IHC.

                          Find your guide

                          Biomarkers of metastasis

                          Validating your biomarker antibody for IHC

                          • Lung cancer biomarkers
                          • Breast cancer biomarkers
                          • Colorectal cancer biomarkers
                          • Prostate cancer biomarkers
                          • Ovarian cancer biomarkers
                          • Pancreatic cancer biomarkers

                          Header image credit: Detecting cancer in human tissues, LM. Credit: Aamir Ahmed, Jane Pendjiky and Michael Millar. Attribution 4.0 International (CC BY 4.0)

                          ​



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