CD20 CD31 CD56 CD45 HMW cytokeratin CAM 5.2 Ki-67 p 53 p 63 HER₂ Vimentin

MELAN- a CK5 CK 16 CYCLIN DI ER PR S100 p504s TTF1 MART-1 KAPPA/LAMBDA

Markers and

AntibodiesLauri Zaker

2015

CD20CD20 is a highly expressed surface antigen that is located mainly on pre-B and mature B lymphocytes.

● Over 95% of B-cell lymphocytes express CD20 as they develop from their pre-B cell stage into plasma cells.

● Unlike other B-cell antigens, CD20 is not shed or internalized upon antibody binding.

o This may allow therapeutic antibodies to recruit immune effector cells and mediate sustained immunologic activity.

● CD20 is believed to play a role in the regulation of calcium transport and B-cell activation and proliferation.

● CD20 is not found on early B-cell progenitors or later mature plasma cells and is not found free in plasma.

http://www.biooncology.com/therapeutic-targets/cd20

http://www.newcomersupply.com/

CD31

http://www.abcam.com/cd31-antibody-ab28364.html

The endothelium of vessels of any tissues are positive.

my IHC.CD31 kidney 40x

A monoclonal antibody against the endothelial cell adhesion molecule CD31.For detection of vascular lesions including benign and malignant vascular neoplasms.Used with Collagen IV sometimes,they together provide a powerful tool for 1) the diagnosis of endothelial neoplasms and 2) the definition of endothelial, mural and pericytic or perivascular tissue compartments in vascular lesions of complex architecture. Hence, we use CD31 and type IV collagen in cases of presumed vascular neoplasms, adding other markers to the panel in accordance with the differential diagnosis, as well as in the recognition of compromised endothelia, such as in vascular invasion by various malignant neoplasms. http://www.ncbi.nlm.nih.gov/pubmed/7479359

CD56 Function This protein is a cell adhesion molecule involved in neuron-neuron adhesion, neurite fasciculation, outgrowth of neurites, etc.

Sequence similarities Contains 2 fibronectin type-III domains.Contains 5 Ig-like C2-type (immunoglobulin-like) domains.Cellular localization Secreted and Cell membrane.

NCAM1 AntibodiesNCAM, as a member of the immunoglobulin superfamily of adhesion molecules is characterized by several immunoglobulin (Ig)-like domains. The extracellular part of NCAM consists of five of these Ig domains and two fibronectin type III homology regions. NCAM is encoded by a single copy gene composed of 26 exons. However, at least 20-30 distinct isoforms can be generated by alternative splicing and by posttranslational modifications, such as sialylation. During sialylation, polysialic acid (PSA) carbohydrates are attached to the extracellular part of NCAM. Through its extracellular region, NCAM mediates homophilic interactions. In addition, NCAM can also undergo heterophilic interactions by binding extracellular matrix components, such as laminin, or other cell adhesion molecules, such as integrins.

Tissue control: pancreas (pos.),lung (neg.)Synonyms CD-56, CD16A, CD16B, CD3 epsilon, CD3-epsilon, CD3e antigen, CD8a, CD8alpha, CD8b, CD8beta, epsilon polypeptide TiT3 complex, epsilon subunit of T3, Fc gamma Receptor 3, Fc gamma Receptor III, FCGR3, FCGR3A, FCGR3B, FCGRIII, FCR-10,FCRIII,FCRIIIA,FLJ18683, Leu-2, MAL, MSK39, NCAM, NCAM1, neural cell adhesion molecule 1,p32,RP11-5K23.1, T-cell antigen receptor complex, T-cell surface antigen T3/Leu-4 epsilon chain, T-cell surface glycoprotein CD3 epsilon chain, T3E, TCRE, CD3e, CD8, NCAM1

http://www.abcam.com/cd46-antibody-epr4014-ab108307.html

ab108307, at 1/500 dilution, staining CD46 in paraffin-embedded Human tonsil tissue

CD46

ab108307 showing positive staining in Thyroid gland carcinoma tissue.

Positive controlMolt-4, Jurkat, HeLa, and K562 cell lysates; Human kidney and Tonsil tissue

ab108307 showing positive staining in breast carcinoma tissue.

ab108307 showing positive staining iin normal breast tissue.

ImmunogenSynthetic peptide conjugated to KLH derived from within residues

1250 to the C-terminus of Human CD45.(Peptide available as ab17550.)Positive control

This antibody gave a positive signal in Jurkat whole cell lysate and Hodgkins lymphoma tissue sections.

http://www.abcam.com/cd45-antibody-ab10559.html

Leucocyte Common Antigen (LCA)

Tissue: (+) Tonsil / (-) Adipose

Function Protein tyrosine-protein phosphatase required for T-cell activation through the antigen receptor. Acts as a positive regulator of T-cell coactivation upon binding to DPP4. The first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Upon T-cell activation, recruits and dephosphorylates SKAP1 and FYN.

Involvement in disease Defects in PTPRC are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)B(+)NK(+) SCID) [MIM:608971].

A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development.

Genetic variations in PTPRC are involved in multiple sclerosis susceptibility (MS) [MIM:126200].

MS is a neurodegenerative disorder characterized by the gradual accumulation of focal plaques of demyelination particularly in the periventricular areas of the brain. Peripheral nerves are not affected. Onset usually in third or fourth decade with intermittent progression over an extended period. The cause is still uncertain.

CD45

This antibody recognizes the HMW (High Molecular Weight) keratin polypeptides of 68, 58, 56.5 and 50 kDa in extract of stratum corneum. The antibody reacts with squamous, ductal and other complex epithelia. It stains adenocarcinomas, breast, pancreas, bile duct, salivary gland and transitional cell carcinomas.

HMW cytokeratin

Cytokeratins are intermediate filament keratins found in the intracytoplasmic cytoskeleton of epithelial tissue. There are two types of Cytokeratins: the low weight, acidic type I cytokeratins and the high weight, basic or neutral type II. Cytokeratins are usually found in pairs comprising a type I Cytokeratin and a type II cytokeratin. The high molecular weight cytokeratins, which are the basic or neutral cytokeratins, comprise subtypes CK1 (67), CK2 (65.5), CK3 (64), CK4 (59), CK5 (58), CK6 (56), CK7 (54), CK8 (52.5) and CK9. The low molecular weight cytokeratins, which are the acidic cytokeratins, comprise subtypes CK10 (56.5), CK12 (56), CK13 (53), CK14 (50), CK16( 48), CK17 (46), CK18 (45), CK19(48) and CK20(46).Cellular localization Cytoplasmic http://www.abcam.com/hmw-cytokeratin-antibody-34be12-ab776.html

\\ staining human skin tissue sections by IHC-P.

human colon carcinoma stained with HMW Cytokeratin, using ABC and AEC chromagen.

CAM 5.2 CloneCAM 5.2Isotype IgG2aImmunogen Anti-Cytokeratin, clone CAM 5.2, is derived

from hybridization of mouse P3/NS-1/1-Ag4-1myeloma cells with spleen cells from BALB/c mice immunized with the human colorectal carcinoma cell line HT29.

Summary Anti-Cytokeratin (CAM 5.2) reagent has a primary reactivity with human keratin proteins that correspond to Moll’s peptides #7 and #8, Mr 48 and 52 kilodaltons (kd), respectively. Cytokeratin 7 and 8 are present on secretory epithelia of normal human tissue but not onstratified squamous epithelium. Anti-Cytokeratin (CAM 5.2) stains most epithelial-derived tissue, including liver, renal tubular epithelium, and hepatocellular and renal cell carcinomas. Anti-Cytokeratin (CAM 5.2) might not react with some squamous cell carcinomas.

Cellular Localization CytoplasmicPositive Control Tissue Lung, colon,

prostate and breast tissue

http://dbiosys.com/products/primary-antibodies/item/5041-cam-52-5041/5041-cam-52-5041

p63 Data supports a role for p63 in squamous and transitional cell carcinomas, as well as certain lymphomas and thymomas.The p63 gene, located on chromosome 3q27-28, is a member of the p53 gene family. The product encoded by the p63 gene has been reported to be essential for normal development.p63 expression is restricted to the nucleus, with a nucleoplasmic pattern. We also observed that the expression was restricted to epithelial cells of stratified epithelia, such as skin, esophagus, exocervix, tonsil, and bladder, and to certain subpopulations of basal cells in glandular structures of prostate and breast, as well as in bronchi.p63 is expressed predominantly in basal cell and squamous cell carcinomas, as well as transitional cell carcinomas, but not in adenocarcinomas, including those of breast and prostate. Interestingly, thymomas expressed high levels of p63. Moreover, a subset of non-Hodgkin’s lymphoma was also found to express p63. Using isoform-specific reverse transcription-PCR, we found that thymomas express all isoforms of p63, whereas the non-Hodgkin’s lymphoma tended to express the transactivation-competent isoforms. We did not detect p63 expression in a variety of endocrine tumors, germ cell neoplasms, or melanomas. Additionally, soft tissue sarcomas were also found to have undetectable p63 levels.

http://clincancerres.aacrjournals.org/content/8/2/494.full

p63

Representative photomicrographs of immunophenotypes of p63 in normal thymus and thymomas, obtained using the anti-p63 4A4 monoclonal antibody. Strong p63 nuclear staining is observed in a population of cells identified as the epithelial elements of the thymus (A). We also observed p63 immunostaining in the neoplastic component of various thymomas, including invasive (B and D), spindle cell (E), and thymic carcinoma (F). Consecutive normal thymus (not shown) and thymoma (B andC) sections were stained with p63 (B) and the anticytokeratin AE1/AE3 monoclonal antibody cocktail (C), revealing that the p63-expressing cells were also costained for cytokeratins and thus are considered of epithelial nature. Original magnifications: ×100 for B and C; ×200 for D and E; ×400 for A and F.

HER2 A protein involved in normal cell growth. It is found on some types of cancer cells, including

breast and ovarian. Cancer cells removed from the body may be tested for the presence of HER2/neu to help decide the best type of treatment. HER2/neu is a type of receptor tyrosine kinase. Also called c-erbB-2, human EGF receptor 2, and human epidermal growth factor receptor 2.

The HER2 gene makes HER2 proteins. HER2 proteins are receptors on breast cells. Normally, HER2 receptors help control how a healthy breast cell grows, divides, and repairs itself. But in about 25% of breast cancers, the HER2 gene doesn't work correctly and makes too many copies of itself (known as HER2 gene amplification). All these extra HER2 genes tell breast cells to make too many HER2 receptors (HER2 protein overexpression)

http://www.gene.com/medical-professionals/medicines/herceptin

It can control the growth of cancer cells that produce too much of a protein called HER2 (human epidermal growth factor receptor

Some breast cancers and stomach cancers have large amounts of HER2 and they are called HER2 positive cancers. HER2 makes the cancer cells grow and divide.

When Herceptin attaches to HER2 it can make the cells stop growing and die.

Trastuzamab a monoclonal antibody

brand name : Herceptin

HER2 for Gastric Cancer Detection

HER2is more than a breast cancer receptor marker; it is also a

Stomach cancer biomarkerOf those diagnosed with the disease, about 22% of people with metastatic

stomach cancer have HER2-positive (human epidermal growth factor receptor-

positive) tumors, which are more aggressive and have a poorer prognosis,

however they might be helped by Trastuzamab.

anti HER2 receptor Trastuzumab for breast, stomachBased on data from a large multicenter phase III trial (ToGA study) trastuzumab has very recently been approved by the EMEA for metastatic gastric cancer and adenocarcinoma of the gastro-esophageal junction. Only patients with tumors which over express Her2 as defined by IHC2+ and a confirmatory FISH+ result, or IHC 3+, determined by an accurate and validated assay are eligible for trastuzumab therapy. However, testing of Her2 status by immunohistochemistry (IHC) differs from breast cancer in core aspects: 1. IHC2+/3+ is scored even though membranous staining is incomplete if membrane staining is clearly detectable even at low magnification (2.5x/5x, 3+) or medium magnification (10x/20x, 2+). 2. Additionally, membrane staining at the appropriate intensity found in at least 10% of tumor cells is restricted to resection specimens. Evaluation of Her2 in situ hybridization (ISH) is similar to breast cancer with ratio values of > or =2.0 indicating Her2 gene amplification. Taking these modifications into account and defining the HER2 positive subgroup as IHC 3+ and IHC2+/FISH+, approximately 16% of gastric cancers are considered Her2 positive, affecting mainly tumor regions with intestinal (gland forming) type carcinoma. In contrast to breast cancer, up to one-third of gastric cancers show a heterogeneous Her2 status both at IHC and ISH levels which favors bright field ISH over FISH.

HER2-CONNECT™: Changing HER2 IHC analysis in important ways

http://www.visiopharm.com/news-and-events--press-releases-page.shtml?page=07-20101117-533595168

Traditional requirements to meticulous outlining of tumor regions

Visiopharm’s patented HER2-CONNECT™ algorithm eliminates the need for manual outlining, leading to time savings

Researchers can work with HER2-CONNECT™ in two different ways: 1) Connectivity and score are immediately computed based on operator defined regions of interest, and 2) Batch processing: Full sections and/or Tissue Micro Arrays can be subjected to analysis in batch processing mode. Computational efficiency makes both approaches possible and feasible on standard desktop computers.

VIMENTINVimentin, a major constituent of the intermediate filament (IF) family of proteins, is ubiquitously expressed in normal mesenchymal cells and is known to maintain cellular integrity and provide resistance against stress. Increased vimentin expression has been reported in various epithelial cancers including prostate cancer, gastrointestinal tumors, CNS tumors, breast cancer, malignant melanoma, lung cancer and other types of cancers. Vimentin's over-expression in cancer correlates well with increased tumor growth, invasion and poor prognosis; however, the role of vimentin in cancer progression remains obscure.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3162105/

Rat cerebral cortex cultures stained with chicken antibody to vimentin ab24525 (green) and rabbit antibody to GFAP (red). Note flattened fibroblastic cells are mostly green (i.e. vimentin positive, GFAP negative), while clearly astrocytic cells, express both vimentin and GFAP and therefore appear golden or orange. Certain other cells express predominantly GFAP and therefore appear red.

Tissue: (+) Melanoma /(-) Adipose

vimentin staining of a tonsilar lymphoma. Note that the epithelium (at the left) is negative.

Melan- A

Melan-A is a melanocyte differentiation antigen, recognized by autologous cytotoxic T lymphocytes. Melan-A is also called MART-1 (melanoma antigen recognized by T cells). The Melan-A/MART-1 gene encodes this protein, 20-22 kDa, associated with endoplasmic reticulum and melanosomes. The function of the protein is unknown. Melan-A is expressed in all normal melanocytes and melanocyte cell lines.Using the monoclonal antibody A-103, staining is also seen in steroid hormone producing cells: adrenal cortex granulosa and theca cells of the ovary and Leydig cells of the testis. This is due to cross reaction (as the Melan-A gene is not detected in these cells).

Fig. 1B. Melan-A staining of normal adrenal cortex using mAb A103. Strong staining of the steroid producing cells. With other melan-A Abs, steroid producing cells are negative.

Fig. 1A. Melan-A staining of normal skin showing strong staining of melanocytes.

Fig. 2A. Melan-A staining of skin with malignant melanoma showing strong staining of normal and neoplastic melanocytes. Novored is here used as chromogene to avoid the merging of melanin and the DAB chromogene.

Fig. 2B. Melan-A staining of renal PEComa (perivascular epitheloid cell tumour)

Fig. 2C. Melan-A (mAb A103) staining of adrenal cortical carcinoma.

Fig. 2D. Melan-A (mAb A103) staining of granulosa cell tumour

http://www.nordiqc.org/Epitopes/melan-a/melan-a-figs.htm

Melan-A

CK5 Western Blot (WB)1:500 - 1:2000Immunofluorescence (IF)1:200 - 1:1000Immunocytochemistry (ICC)1:200 - 1:1000Immunohistochemistry (IHC)1:200 - 1:1000Flow Cytometry (FACS)1:200 - 1:400* Suggested working dilution

http://www.pierce-antibodies.com/CK5-antibody-clone-2C2-Monoclonal--MA517057.html#

The protein encoded by this gene is a member of the keratin gene family. The type II cytokeratins consist of basic or neutral proteins which are

arranged in pairs of heterotypic keratin chains coexpressed during differentiation of simple and stratified epithelial tissues.

This type II cytokeratin is specifically expressed in the basal layer of the epidermis with family member KRT14.

Mutations in these genes have been associated with a complex of diseases termed epidermolysis bullosa simplex. The type II cytokeratins are clustered in a region of chromosome 12q12-q13.

control: squamous/stratified squamous

CK16 Positive controlHuman breast carcinoma tissue and HepG2 cell extracts.

Mouse monoclonal to CK16. Keratin 16 is expressed in keratinocytes, which are undergoing rapid turnover in the suprabasal region ,also known as hyperproliferationrelated keratins). Keratin 16 is absent in normal breast tissue and in noninvasive breast carcinomas. Only 10% of the invasive breast carcinomas show diffuse or focal positivity. Reportedly, a relatively high concordance was found between the carcinomas immunostaining with the basal cell and the hyperproliferationrelated keratins, but not between these markers and the proliferation marker Ki67. This supports the conclusion that basal cells in breast cancer may show extensive proliferation, and that absence of Ki67 staining does not mean that (tumor) cells are not proliferating.

http://www.biorbyt.com/ck16-antibody-3

human gullet cancer tissue using CK16

antibody

CYCLIN D1 antibodyThe protein encoded by this gene belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance throughout the cell cycle. Cyclins function as regulators of CDK kinases. Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. This cyclin forms a complex with and functions as a regulatory subunit of CDK4 or CDK6, whose activity is required for cell cycle G1/S transition. This protein has been shown to interact with tumor suppressor protein Rb and the expression of this gene is regulated positively by Rb. Mutations, amplification and overexpression of this gene, which alters cell cycle progression, are observed frequently in a variety of tumors and may contribute to tumorigenesis.(RefSeqJul2008)

Aliases of CCND1Cyclin D1 PRAD1 OncogeneBCL1 Cyclin D1 (PRAD1: Parathyroid Adenomatosis 1)

PRAD1 G1/S-Specific Cyclin D1D11S287E Parathyroid Adenomatosis 1B-Cell CLL/Lymphoma 1 U21B31B-Cell Lymphoma 1 Protein G1/S-Specific Cyclin-D1 BCL-1 Oncogene PRAD1 Oncogene

Immunogen Synthetic peptide corresponding to Human

Cyclin D1 (C terminal).

EpitopeC-terminus

Positive control Breast carcinomas, mantle cell lymphoma,

MCF7 cell lysate

http://www.abcam.com/cyclin-d1-antibody-sp4-ab16663.html#description_images_1

mouse testis tissue, staining Cyclin D1 with ab16663.

http://www.genecards.org/

EREstrogen Receptor Alpha (ER Alpha) is a nuclear protein and member of the steroid hormone receptor family. ER alpha possesses both DNA binding and ligand binding domains, and exerts a significant role in activating the transcription of certain genes. Ligand-dependent dimerization and phosphorylation both function to regulate the transcriptional activation of ER alpha.

http://www.epitomics.com/diagnostics/product/2156

PRPresent in hormone responsive tissue and their neoplasm. also reported in carcinoma of lung,stomach and thyroid. STUMP (smooth muscle tumor of uncertain malignant

potential )and stromal sarcomas are positive for PR. Handbook of Practical Immunohistochemistry: Frequently Asked Questions edited by Fan Lin, Jeffrey Prichard

PR y85

cellmarque.com/antibodies/Progesterone-Receptor-Y85 cellmarque.com/antibodies/Progesterone-Receptor-SP42

PR SP42

PR

Formalin-fixed, paraffin-embedded human breast carcinoma stained with PR antibody using peroxidase-conjugate and AEC chromogen. Note nuclear staining of tumor cells.

https://www.lifetechnologies.com/order/genome-database/antibody/Progesterone-Receptor-Antibody-hPRa-2-Monoclonal/MA5-12642

S100Small dimeric member of the family of calcium-binding proteins.Widely distributed in human tissues, including glia, neurons,chondrocytes, Schwann cells,melanocytes,phagocytic or antigen-presenting mononuclear cells. Langerhans,histiocytes,myoepithelial cells and various epithelia.

http://www.abcam.com/s100-beta-antibody-astrocyte-marker-ab41548.html

ab41548 staining S100 beta in mouse brain tissue sections by Immunohistochemistry (IHC-P -

paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde

and blocked with 1% BSA for 10 minutes; antigen retrieval was by heat mediation in

citric acid. Samples were incubated with primary antibody (1/10000 in TBS/BSA/azide)

for 2 hours at 21°C. A Biotin-conjugated goat anti-rabbit IgG polyclonal (1/250) was

used as the secondary antibody.

At this dilution factor, there is very good delineation of astrocyte processes.

p504s

Alpha-methylacyl-CoA racemase (AMACR), also known as p504s, is a mitochondrial and peroxisomal

enzyme that is involved in bile acid biosynthesis and beta-oxidation of branched-chain fatty acids. AMACR is

essential in lipid metabolism, and is expressed in normal liver (hepatocytes), kidney (tubular epithelial cells)

and gallbladder (epithelial cells). Expression has also been found in lung (bronchial epithelial cells) and colon

(colonic surface epithelium). Expression is granular and cytoplasmic. AMACR expression can also be found

in hepatocellular carcinoma and kidney carcinoma. Past studies have also shown that AMACR is expressed

in various colon carcinomas (well, moderately and poorly differentiated) and over expressed in prostate

carcinoma.

Human prostatic adenocarcinoma: immunohistochemical staining for alpha-methylacyl-CoA racemase (AMACR, p504S) using NCL-L-AMACR. Paraffin section.

http://www.leicabiosystems.com/ihc-ish-fish/novocastra-reagents/primary-antibodies/details/product/alpha-methylacyl-coa-racemase-amacr-p504s/

TTF-1This gene encodes a transcription termination factor that is localized to the nucleolus and plays a critical role in ribosomal gene transcription. The encoded protein mediates the termination of RNA polymerase I transcription by binding to Sal box terminator elements downstream of pre-rRNA coding regions. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. This gene shares the symbol/alias 'TFF1' with another gene, NK2 homeobox 1, also known as thyroid transcription factor 1, which plays a role in the regulation of thyroid-specific gene expression. [provided by RefSeq, Apr 2011]

In lung adenocarcinomas, positive and partial positive TTF-1 expression has a significant positive correlation with EGFR mutations(exon 19 and 21). In clinical practice, TTF-1 expression combine with EGFR mutations, especially exon 21 mutation can guide clinical treatment timely for lung adenocarcinomas .

Citation: Shanzhi W, Yiping H, Ling H, Jianming Z, Qiang L (2014) The Relationship between TTF-1 Expression and EGFR Mutations in Lung Adenocarcinomas. PLoS ONE 9(4): e95479. doi:10.1371/journal.pone.0095479

TTF-1 positive expression in adenocarcinoma cell (IHC×400).

transcription termination factor, RNA polymerase I

MART-1 A tumor-associated melanocytic differentiation antigen. Vaccination with MART-1 antigen may stimulate a host cytotoxic T-cell response against tumor cells expressing the melanocytic differentiation antigen, resulting in tumor cell lysis.

Synonyms:Antigen LB39-AAAntigen SK29-AAMART-1MART-1 Tumor AntigenMelan AMelan-A ProteinMelanoma Antigen Recognized by T-Cells 1MLANAMLANA Protein

● Commonly used melanocytic markers; MelanA also called A103; are separate antibodies that occasionally have different staining properties● Melanocytic Antigen Recognized by cytotoxic T lymphocytes from melanoma patients● Cytoplasmic protein sensitive and specific for melanoma and melanocytic lesions

Uses by pathologists=========================================================================● Recommended for evaluating sentinel lymph nodes for melanoma (Am J Surg Pathol 2001;25:1039), although also stains benign nevi (Am J Surg Pathol 2002;26:1351) and normal melanocytes● Differentiate adrenal cortical tumors (MART1+) vs. renal cell carcinoma or pheochromocytoma (MART1-)● Differentiate neurotized melanocytic nevi (MelanA+) from neurofibroma (MelanA-, Arch Pathol Lab Med 2012;136:810)● Distinguish in situ and invasive melanoma, and measure thickness (Am J Dermatopathol 2012 Jun 8 [Epub ahead of print])● Caution: may overestimate presence of lentigo maligna (Dermatol Surg 2011;37:657, J Cutan Pathol 2011;38:775)

MART-1 mRNA (blue/purple color) is seen by RT in situ PCR

in melanophages with coarse pigment granules (Panels A&B).

Immunohistochemistry for MART-1 shows a weakly positive

signal in scattered large cells with “normal” nuclei and coarse

melanin granules, consistent with melanophages (Panel C).

Immunohistochemistry for HMB-45 shows a positive signal in

scattered large cells, consistent with melanophages (Panel D).

Double staining of MART-1 mRNA (blue/purple color) and

CD68 antigen (red color) shows that some cells co-express

both MART-1 mRNA and CD68 (arrow), while MART-1

mRNA and CD68 are separately expressed by other cells.

(Panels E&F). Sentinel node (Panel G) and NSN (Panel H)

from a breast cancer patient are negative for MART-1 mRNA.

Am J Surg Pathol. Author manuscript; available in PMC 2012 Nov 1.Am J Surg Pathol. 2011 Nov; 35(11): 1657–1665.

doi: 10.1097/PAS.0b013e3182322cf7

KAPPA/LAMBDAImmunohistochemical staining for kappa and lambda light chains is often performed on lymphoid tissue. Staining for kappa and lambda light chain expression is notoriously problematic—some might say finicky. One complication is the need for decalcification on some lymphoid tissue often stained for kappa and lambda, such as bone marrow core biopsies. The more common fixatives currently in use have different effects on the various tissues under study and may impart different properties to the protein targets in question.

Anti-kappa and anti-lambda detect surface light chain immunoglobulins on normal and neoplastic B-cells in human lymphoid tissue. In normal lymphoid tissue the kappa and lambda cell ratio is approximately 2:1, but values in excess of that ratio indicate monoclonality caused by either a lymphoproliferative disorder or neoplasia such as lymphoma.

Both kappa (M) (brown) and lambda (P) (red) seen on the same tissue section, thus allowing the end-user a more accurate and easier assessment of both stains. Image analysis of a digitally scanned slide provides rapid highly accurate kappa and lambda cell counts. The resulting whole slide section determination of a kappa/lambda ratio enables easier diagnosis of disease.

http://flagshipbio.com/lymph/kappalambda-in-lymphoma/