IIImmunohistochemistry (IHC) Outreach Services

 Note type of fixative used if not neutral buffered formalin.  Note type of tissue/specimen  Unless specified otherwise, positive and negative controls react satisfactorily.

Available Chromogen – All markers have been validated with 3,3’-Diaminobenzidine Tetrahydrochloride (DAB) which results in a brown/black precipitate. DAB is the routine chromogen. In addition, some markers have also been validated using the Fast Red (RED), which results in a red precipitate. If available with both chromogens and one is not selected, the def ault will be the DAB chromogen.

Antibody Common Applications St aining Charac teristi cs

Actin (muscl e sp eci fic ) Smoo th, ske letal & ca rdiac musc le Cytoplasm ic Actin (smoo th muscle ) Smoo th muscl e an d myoep itheli al cel ls Cytoplasm ic and memb rane ALK Pr otein ALK1 posi tive lymp homa s Cytoplasm ic and/or nuclea r Alph a-1-Antitr ypsi n Demo nstr ates A-1-AT in liver Cytoplasm ic (A-1-AT) Bcl-2 Oncop rotein Foll icula r lymph oma an d so ft tiss ue Cytoplasm ic tumors Bcl-6 Foll icula r lymph oma Nucl ear Ber-EP 4, Epithelia l Antige n Aden oca rcin oma vs. meso the liom a Memb rane and cytoplasm ic. The and epithelial tumors membrane staining is preferentially basolateral. Beta-Amyloid Pos t mo rt em dia gnos is of demen tia E xtr acel lular de pos ition (am yloid plaques), vascular deposition (amyloid angiopathy) BRS T-3, (B72.3) Aden oca rcin oma vs. meso the liom a Cel l su rfac e an d cytop la sm ic Beta -Catenin Desmoid type-fibromatosis, solid and Nuclear pseudopapillary tumors of pancreas . CD 1a Langerhan cells, thymic T-cells, Memb rane and weak ly cytoplasm ic thymoma CD3 T-cells , lymp hom a/leuk emi a typin g Memb rane CD4 Helper/Inducer T-cells mycosis Memb rane fungoides vs. cutaneous inflammatory processes, / leukemia typing CD5 T-cell , lymphoma /le ukemi a typin g Memb rane CD8 T-cel l supp resso r/ cytoto xic , Memb rane lymphoma/leukemia typing, mycosis fungoides vs. cutaneous inflammatory processes CD 10 Lymph oma typing , me tas tatic Cytoplasm ic and Memb rane carcinoma unknown primary CD 15 Hodgkin Lymphoma Typing, Reed -St ernbe rg cell s in Hodgkin’ s mesothelioma vs. adenocarcinoma lymphoma show cell membrane and granular paranuclear staining. CD 20 , B C el l Lymph oma /le ukem ia typing Cytoplasmic side of the cell surface membrane CD21 Follicular Dendritic cell marker, found Cytoplasmic & Membrane also on some B cells CD 23 Lymph oma /le ukem ia typing Cytoplasm ic/memb ran e

Antibody Common Applications St aining Charac teristi cs

Revised 12/4/19 Page 1 of 6 L. Bliven CD 30 Anapl astic la rge cel l lymph oma, Memb rane and/or a dot like cytopl asmi c Hodgkin lymphoma staining CD 31 Endo thel ial Cells Pr edomina tel y cell memb rane , with weaker cytoplasmic staining. CD 34 Soft tissu e tumo r cla ssi fica tio n, Memb rane/cytoplasm ic leukemia typing CD 43 T-cell , lymphoma /le ukemi a typin g Pr edomin antly confine d to the cel l surface CD 45 (LCA) Lympho hema topoe tic tumo rs Memb rane, but cytoplasm ic may als o occur CD 56 NK cel ls, tum ors de rived from Memb rane neuroectodermal tumors such as CD 57 Lneuymphroeondmoac tryinpeing an, dneu neruorenobdlaosc tromin ea s, Cytoplasm ic and memb ran e differentiation CD 68 , PG- M1, Mac rop hag e His tiocytic /monoc ytic ma rke r Cell s of mon ocyte/mac roph ag e li nea ge stain diffuse or granular cytoplasmic. Mast CD 79 a B-cel l, lymphoma /le ukem ia typin g Ccelellls Maembre neragnaeti vaen. d/or cytopl asm

CD 99 , SEE MIC2 CD 11 7 (c-ki t) Gas tr ointes tinal Str omal Tum ors Memb rane and/or cytopl asmi c (GIST), Mast Cells, Stains approximately 75% of mesenteric fibromatosis tumors. CD 13 8 Synd eca n-1 Plasm a Cell s (also stains en dothe lia l Cel l Memb rane, pre- an d pl asm a cells, fibroblasts, keratinocytes, and cell marker, but is absent from mature B CD 16 3 Hno is rtmioacly htiecpa Letocsi oyntess ) Mcelembls. Irta ins ea se lec tive ma rke r for B ce ll

CDX2 Colo n an d othe r GI ca nce rs are str on g Nucl eus in no rmal and neop lastic & diffusely positive. Also, mucinous intestinal epithelial cells. Calci ton in Moveadrulia lna rcay Tnhceyros ida reC aprocisintomivea. Neg for Cytoplasm ic

Cal retini n Meso theli oma vs. ad enoca rci n oma , Cytoplasm ic and nucl ear sex cord stromal, adrenal tum ors, & Carci noemb ryon ic Antigen , CEA/M MHesoirsc hthperuli onmg’ as vcsa.s Aesde. Pnocleaasrec isnpecomaific, y Cytoplasm ic. Norma l co lon-CEA is metastatic carcinoma of unknown mainly localized at the apical border of the Carci noemb ryon ic Antigen , CEA/P Hprepaima tryom a vs. ad enoca rcinoma Cepyittoplasmheli al celic.l sIn. C li ovelo r,n pcarerdcoinomminana- tly bilary canaliculi are labeled. In colonic Chrom ogran in A Neu roe ndoc rin e di fferen tiation Cadenytoplasmoca rciinc om a the stain ing reac tio n is

Cycli n D1 Man tle cel l lymphom a Nucl ear

Cytoke ratin 5/6 Meso theli oma vs. Ade noc arcinom a Cytoplasm ic

Cytoke ratin 7 Metastatic carcinoma of unknown Cytoplasmic. E xceptions to reactivity may primary occur e.g. CK-7 positive hepatocytes have been observed in patients with acute and chronic cholestasis. Cytok eratin 20 Metastatic carcinoma of unknown Cytoplasm ic. May occasio nall y be primary e xpressed in breast and lung adenocarcinomas, and in squamous cell carcinomas. Less than 5% CK20 positive cells may be present in a number of tissues not generally considered CK 20 positive.

CK 11 6 – MNF 11 6 Epi theli al ma rke r Cytoplasm ic Cytoke ratin 34 BE12 Prostate basal cells, squamous cells Cytoplasm ic vs. adenocarcinoma Cytoke ratin , AE1/AE3 Epi theli al tumo rs, hepa tom a vs. Cytoplasm ic adenocarcinoma Cytoke ratin , CA M5.2 Epi theli al tumo rs Cytoplasm ic

Revised 12/4/19 Page 2 of 6 L. Bliven Antibody Common Applications St aining Charac teristi cs

Cytoke ratin Cock tail (K erCK) Epi theli al tumo rs Cytoplasm ic Cytomega lovi rus /IHC Cytomega lovi rus Nucl ear stai nin g pa ttern in ea rly HC MV infection, later stage, cytoplasmic staining might be observed. Does not x- react with adenovirus, herpes simplex virus, & varicella zoster virus. D2-40 Lympha tic end otheli um, Cytoplasm ic and some tim es memb rane mesothelioma vs. adenocarcinoma Desm in Smoo th and ske letal musc le Cytoplasm ic, may sh ow a fibrilla ry differentiation aspect. Eps tein -Barr Virus (EBV) RNA CISH La ten t EBV in fec tio n Nucl ear. Surgical /H ema topa th olog y consultation and review of the entire case is highly recommended. E-Cadhe rin Lobu lar vs. duc tal breast ca rcinom a Cellu lar mem bran e, some cytoplasm ic

Epi theli al Memb ran e Antigen (E MA) Metas tatic ca rci nom a of unk nown In ne opl asms , cytop la sm ic and ap ica l primary, lymphoma luminal membrane staining are the most common patterns of immunoreactivity with peripheral membrane staining or other patterns also occurring. Plasma cells stain positive. In normal breast and other secretory epithelia, labeling is predominantly localized to apical luminal membranes.

Estr ogen Recep tor (ERA) Br eas t ca rci nom a prognostic ma rke r, Nucl ear, cytopl asmi c is co nsi dered no n- metastatic carcinoma of unknown specific Occasional lymphoid tumors and primary non-lymphoid neoplasms such as melanomas are labeled. Follow CAP & ASCO guidelines. Fac tor VIII Rela ted Ag (Von Megaka ryoc ytic an d endo the lial Cytoplas m as diffu se or some tim es W illebrand Factor) marker slightly granular staining

GATA-3 Ur ot heli al ca rci noma , breast ducta l Mus t be nuc le ar, stron g or mode rate epithelium, and transitional cells intensity, and non-focal in urothelial carcinoma.

Glia l Fib rilla ry Acid ic Pr otei n GFAP Glia l tumo rs Cytoplasm ic Aceton e fixed fro zen or Bouins fixed tissues label certain neuronal structures, including a xons, indicting a X-r xn w/ neurofilam ent -3 Hepatocellular carcinoma vs. benign Granula r cytop lasm ic, and memb ranous hepatocellular lesions Patterns Yolk Sac Tumor, Choriocarcinoma

Helicob acter P ylo ri Helicob acter pylo ri in fection Indi vid ual H. pylo ri bac terium when present on the surface of the epithelium or in the cytoplasm of the epithelial cells. ASR

Hemo glob in A Er ythoi d ma rker Cytoplasm ic

Revised 12/4/19 Page 3 of 6 L. Bliven Antibody Common Applications St aining Charac teristi cs

Hep Par1 (H ep Ab) Hepa toma , gas tric ca rci nom a Disp lays a dis tinct, gran ula r cytopl asmi c staining pattern, which is occasionally ring- like and is present diffusely throughout the hepatocyte cytoplasm HER 2/NEU c -erbB -2 Oncoprotein HER-2/neu overexpression for invasive Follo w CAP an d ASCO guid elines for breast and gastric cancers. interpretation. Laboratory Developed Test (LDT).

Herpes Simplex Virus I (HSVI) HSV Infection Nuclear and cytoplasmic

Herpe s Sim ple x Virus II (HSV II ) HSV Infection Nucl ear and cytoplasm ic

HMB-45 , Melanos ome Mela noc yte ma rke r, angiomyolipom a Cytoplasm ic. Orde r DAB or RED hMLH-1 Scree nin g for Lynch Synd rome Nuclear – stains normal cells and non- mutant cells hMSH -2 Scree nin g for Lynch Synd rome Nuclear – stains normal cells and non- mutant cells hMSH -6 Scree nin g for Lynch Synd rome Nuclear – stains normal cells and non- mutant cells IgG Plasm a Cell Marke r, use d in ratio with Cytoplasm ic IgG4 for IgG4 related diseases IgG4 Plasm a Cell Marke r, use d in ratio with Cytoplasm ic IgG for IgG4 related diseases Inhi bin , Alpha Adrena l co rtical , sex-co rd str omal Cytoplasm ic tumors Ins uli n Panc reatic endoc rin e tumo rs Cytoplasm ic

Kapp a Lig ht Chains Plasmac ytomas, neop las ti c lymph oid Cel l memb ran e an d/or cytoplas m tumors vs. reactive proliferations

Kappa /Lambd a Doub le or Singl e Demo nstr ate clon ali ty in leuk emi as, Comb ina tion of Kap pa (RED) and Stain RNA CISH plasmacytomas, and certain non- Lambda (DAB). Cytoplasmic. Hodgkin

Ki-67 Cell proliferation marker Nuclear, except in mitotic cells where the and cytoplasm are labe led Occasional labeling of tissue components in vessel walls and KiMart Ki-67 and Melan A. See separate Ki-67 nuclear DAB and Melan A antibodies. cytoplasmic RED. Lam bda Ligh t Cha ins Plasmacytomas, neoplastic lymphoid Cel l memb ran e an d/or cytoplas m tumors vs. reactive proliferations

Lu teini zin g Hormone , LH Pitui tary neo plasm s Cytoplasm ic

Mel an-A (A 103) Melanocyte marker, adrenal cortical, Cytoplasmic The Melan-A is also sex-cord stromal tumors called MART-1. Order DAB or RED

Mela nom a Cock tai l (Mel CK) Mela noc yte ma rker Cytoplasmic. Combination of HMB-45, two clones of MART-1 and Tyrosinanse antibodies. Order DAB or RED.

Antibody Common Applications St aining Charac teristi cs

Revised 12/4/19 Page 4 of 6 L. Bliven MIC 2, CD99 PNE T/Ewin gs sa rcoma Cel l memb rane and Cytoplasmic Mic rophtha lmi a Transc rip tio n Fac tor Mela noc yte ma rker Nucl ear, cytopl asmi c is no t consi dered (MITF) positive, Order DAB or RED. MOC-31 Epi theli al related an tig en Memb rane

MUM1 Germin al cen ter B cel ls, activated T Nucl ear posi tivity with wea k to mode rate cells, plasma cells, and melanocytes cytoplasmic staining

MYC ( c-MyC) Diff us e La rge B-cel l lymphomas Posi tive is a nuc lea r stain in >40 % of (DLBCL) cells staining moderate to strong. Myel operoxidas e Myel oid ma rker Cytoplasm ic Occas ion al stai ning of histiocytes was observed, possibly due to phagocytosed material MyoD1 Rhabd omyosa rcoma Nuclear. Results of a study suggest that expression in rhabdomyosarcomas is inversely related to the degree of cellular differentiation of the tumor cells. Only Myogenin Rhabd omyosa rcoma Nuclear. Nuclear e xpression has been reported to be inversely related to the degree of cellular differentiation of rhabdomyosarcoma tumor cells Myog lobi n Rhabd omyosa rcoma Cytoplasm ic Naps in A Pulm ona ry Ade noca rcinom a Cytoplasm ic Neu rofilament Neu ral tumo rs & neu rons Cytoplasm ic

Neu ron Speci fic Enolase NSE Can be found in virtually any type of Cytoplasmic. Neurons are labeled in both neoplasm cytoplasm and processes. Oct-3/4 Classic Seminoma/Dysgerminoma, Strong nuclear reactivity, with cytoplasmic Embryonal Carcinoma, staining Gonadoblastoma, and Intratubular Germ Cell Neoplasias p16 Cervica l dyspl asia Both nuclear and cytoplasmic. Look for diffuse and intense staining. Focal and sporadic is considered negative. p40 Pulmonary squamous cell carcinomas, Nuclear more specific than p63. p5 3 Pr otein p5 3 tumo r supp resso r ge ne produc t Usua lly nuclea r, bu t cytoplas mic has been reported in some cases Sections must be cut fresh, day of staining. p5 7 For classi fic atio n of mo lar pregn anc y Nuclear stain in cytotrophoblasts and decidual proliferating basal cells of cervix cells. Hydropic abortus and partial moles=positive stromal cells. Complete moles=negative stromal cells. P63 Antibody Basal cells in the prostate gland, Nucl ear myoepithelial cells in breast, p63 /CK 5/CK 14 Stains basal cells of normal and p63 nuclear DAB, CK5/CK14 benign prostate glands, and cytoplasmic DAB. myoepithelial cells of breast. p504S, Alpha-Methylacyl- Prostate adenocarcinoma and atypical Granular/Cytoplasmic Best if used in CoA Racemace (AMACR) adenomatous hyperplasia conjunction with high molecular weight cytokeratin. Order DAB or RED. PA X-5 B-cel l, lymphoma /leukem ia typing Nucl ear PAX -8 Ovarian serous ca, renal cell ca, and Nuclear thyroid tumors Antibody Common Applications St aining Charac teristi cs

Revised 12/4/19 Page 5 of 6 L. Bliven PDL -1 NSCLC, GEJ adenoca, urothelial Cell membrane of tumor cells & cell membrane carcinoma, & other tumors/immune cells and/or cytoplasm of immune cells PIN4 See sepa rate ma rke rs p6 3 nuc lea r DAB, CK 5/CK 14 cytoplasm ic DAB, & P504S RED. PMS2 Scree nin g for Lynch Synd rome Nuclear – stains normal cells and non-mutant cells Pr oges terone Recep tor, PRA Br eas t prognos tic ma rker Nucl ear, cytopl asmi c is co nsi dered non - specific. Follow CAP & ASCO guidelines. Prolactin Pituitary neoplasms Cytoplasmic Pr os tate Cock tail Pr os tate ca rcinoma Comb o of PAP and PSA A ntibodi es

Pr os tate Speci fic Antigen PSA Pr os tate ca rcinoma Cytoplasm ic. St ain ing is pred omin antly intracytoplasmic and secretions are also frequently stained positively.

Pr os tatic Aci d Phosp has tase (PAP) Pr os tate ca rcinoma Cytoplasm ic Prostein (P501S) Expressed in both benign and neoplastic prostate Cytoplasmc tissue -Prostate Carcinoma S100 Mela noma , neu ral ma rker Cytoplasm ic. Orde r DAB or RED.

Smoo th Muscl e Myosi n Heavy Chai n Myoep ithe lial ma rke r, smoo th muscl e Cytoplasm ic (SMMHC) differentiation Synap toph ysi n Neu roe ndoc rin e di fferen tiation Cytoplasm ic pa ttern, occ asio nall y revealing a punctuate or granular pattern Synucle in (alpha) Pos tmo rt em diag nos is of Parkinso n’s Cytoplasmic (Lewy bodies, Lewy Disease and Lewy body dementia. neurites). Tau Postmortem diagnosis of Alzheimer’s Cytoplasm ic (neurofi brilla ry tangl es, Pick disease and frontotemporal dementia. Bodies). Termi nal Deoxynucleo tidyl Ele vated le vels ha ve bee n report ed in Nucl ear Transferase (TdT) tumor cells of lymphoblastic lymphoma (MLLB), lymphoid blast crisis of chroni myeloid leukemias Thyroglobul in Thyroid ca rcin omas St ain ing is con fin ed to the lumen of thyroi d follicles and the apical surface of thyrocytes. In carcinomas it may also display staining of the thyrocyte cytoplasm Tryptas e Mas t cel ls Granula r cytop lasm ic stai nin g pa tt ern, corresponding to the secretory granules of mast cells

TTF-1, Thyroi d Transc ription Fac tor Lung & thyroid marker, also some Nucl ear 1 neuorendocrine Tyrosi nas e Mela noc ytic lesio ns Cytoplasm ic. Orde r DAB or RED.

Vime ntin Metastatic carcinoma of unknown Cytoplasm ic primary, sarcomas W il ms ’ Tumo r (W T1) W il ms ’ Tumo r, se rous ca rcin oma , & Nucl ear other tumors

Revised 12/4/19 Page 6 of 6 L. Bliven