Method/Kit:
Immunocytochemistry and immunofluorescence protocol

Measurement Type Imaging
Description Immunocytochemistry (ICC) combines anatomical, immunological and biochemical techniques to image discrete components in tissues by using appropriately-labeled antibodies to bind specifically to their target antigens in situ.

Target/Analytes with this Method/Kit
Target/Analyte Short Name Description
Actin cytoskeleton | Mitochondria | Nucleus Actin-mito-nuclei Phalloidin stained actin cytoskeleton; ATPB containing mitochondria; DAPI stained nuclei
Aquaporin 1 AQP1 Aquaporin 1
Bile canaliculi | Nucleus Bile canaliculi MRP2-containing bile canaliculi DAPI stained nuclei
CD31 CD31 CD31, also known as platelet endothelial cell adhesion molecule-1 (PECAM-1), is a transmembrane homophilic receptor that is expressed by endothelial cells (ECs), platelets, granulocytes, macrophages, dendritic cells (DCs), T- and B-cells and natural killer (NK) cells. As a member of the immunoglobulin (Ig) gene superfamily, CD31 comprises six extracellular Ig folds; it has a molecular mass of 130 kDa and is differentially glycosylated, involving N-linked and O-linked glycosylation sites. [Federica M. Marelli-Berg, Marc Clement, Claudio Mauro, Giuseppina Caligiuri Journal of Cell Science 2013 126: 2343-2352; doi: 10.1242/jcs.124099 ]
CD31 | Desmin | Nucleus CD31 | Desmin | Nucl CD31 | Desmin | Nucleus
CD68 | Desmin | Nucleus CD68 | Desmin | Nucl CD68 | Desmin | Nucleus
COL Ia1 COL Ia1 Fluorescent Expression of the COL1A1 Protein
Cytokeratin 8/18 CK 8/18 Human Hepatocyte specific cytokeratin intermediate filaments
FITC-Dextran FITCdex Direct read of FITC-Dextran in media effluent typically to assess permeability.
GABAergic and Glutamatergic neurons GABA/Glutamatergic Quantification of both GABAergic and Glutamatergic neurons in the preBotC neuron population
GABAergic neurons GABAergic Quantification of GABAergic neurons in the preBotC neuron population
Glutamatergic neurons Glutamatergic Quantification of glutamatergic neurons in the preBotC neuron population
HO-1 in Cells HO-1 Immunocytochemistry imaging.
Homeobox A1 (HOX A1) HOXA1 HomeoboxA1
Homeobox B1 (HOX B1) HOX B2 Homeobox B2
Homeobox D1 (HOX D1) HOX D1 Homeobox D1
Image IMG Image for later analysis of intensity, morphology, distribution, location, etc.
OAT1 OAT1 Oat1 [Slc22a6, originally identified as NKT (novel kidney transporter)] plays a central role in the metabolism and handling of gut microbiome metabolites as well as putative uremic toxins of kidney disease. The organic anion transporter (OAT) subfamily has a role in handling of common drugs (antibiotics, antivirals, diuretics, nonsteroidal anti-inflammatory drugs), toxins (mercury, aristolochic acid), and nutrients (vitamins, flavonoids). [Nigam SK, Bush KT, Martovetsky G, et al. The organic anion transporter (OAT) family: a systems biology perspective. Physiol Rev. 2015;95(1):83-123. doi:10.1152/physrev.00025.2013]
OCT-2 Quantification (Imaging) OCT-2 Image based quantification of OCT-2 staining
octamer-binding transcription factor 4 Oct-4 Oct-4 is a homeodomain transcription factor of the POU family. It is critically involved in the self-renewal of undifferentiated embryonic stem cells.
SGLT2 SGLT2 Glucose transporter
Steatosis STEATOSIS Abnormal retention of lipids.
VE-Cadherin VE-Cadherin VE-cadherin belongs to the super-family of classical cadherins, and as such, mediates homotypic calcium-dependent cell–cell interactions.1 VE-cadherin expression is tissue-specific and exclusive to endothelial cells. Also know as Cadherin 5, type 2, or CD144 (Cluster of Differentiation 144). [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4049861/]
ZO-1 ZO-1 Recommended name: Tight junction protein ZO-1 Alternative name(s): Tight junction protein 1 Zona occludens protein 1 Zonula occludens protein 1 TJP1, TJP2, and TJP3 are closely related scaffolding proteins that link tight junction (TJ) transmembrane proteins such as claudins, junctional adhesion molecules, and occludin to the actin cytoskeleton (PubMed:7798316, PubMed:9792688). [https://www.uniprot.org/uniprot/Q07157]
Actin cytoskeleton | Mitochondria | Nucleus Actin-mito-nuclei Phalloidin stained actin cytoskeleton; ATPB containing mitochondria; DAPI stained nuclei
Albumin ALB Albumin is the major constituent of serum protein (usually over 50%). It is manufactured by the liver from the amino acids taken through the diet. It helps in osmotic pressure regulation, nutrient transport and waste removal. High levels are seen rarely in liver disease, shock, dehydration, or multiple myeloma. Lower levels are seen in poor diets, diarrhea, fever, infection, liver disease, inadequate iron intake, third-degree burns and edemas or hypocalcemia. Normal Adult Range: 3.2 - 5.0 g/dl Optimal Adult Reading: 4.1
Bile canaliculi | Nucleus Bile canaliculi MRP2-containing bile canaliculi DAPI stained nuclei
BSEP Protein BSEP BSEP protein levels measured by immunoflourescence
CD31 CD31 CD31, also known as platelet endothelial cell adhesion molecule-1 (PECAM-1), is a transmembrane homophilic receptor that is expressed by endothelial cells (ECs), platelets, granulocytes, macrophages, dendritic cells (DCs), T- and B-cells and natural killer (NK) cells. As a member of the immunoglobulin (Ig) gene superfamily, CD31 comprises six extracellular Ig folds; it has a molecular mass of 130 kDa and is differentially glycosylated, involving N-linked and O-linked glycosylation sites. [Federica M. Marelli-Berg, Marc Clement, Claudio Mauro, Giuseppina Caligiuri Journal of Cell Science 2013 126: 2343-2352; doi: 10.1242/jcs.124099 ]
CD31 | Desmin | Nucleus CD31 | Desmin | Nucl CD31 | Desmin | Nucleus
CD68 | Desmin | Nucleus CD68 | Desmin | Nucl CD68 | Desmin | Nucleus
Cell Morphology Cell Morphology Imaging, typically with transmitted light, to see the cell morphology.
Cell Morphology & Confluency Morphology Plus Cell Morphology & Confluency
Collagen IV a1,2 Col4A1,2 Collagen type IV alpha 1 and 2
Collagen IV a1,2 | DNA | Laminin Col4A1,2|DNA|Laminin Multi-stain imaging target
Collagen IV a4 Col4A4 Collagen, type IV, alpha 4
CYP3A4 CYP3A4 Cytochrome P450 3A4 (abbreviated CYP3A4) is an important enzyme in the body, mainly found in the liver and in the intestine. It oxidizes small foreign organic molecules, such as toxins or drugs, so that they can be removed from the body. [https://en.wikipedia.org/wiki/CYP3A4]
Cytokeratin-18 CK-18 EMT marker.
Cytokeratin 19 CK-19 Cytokeratin 19
Dead Cells DEAD Number or percent of dead cells
DNA DNA Immunocytochemistry imaging.
DNA | Albumin | ABCB11/BSEP DNA/ALB/BSEP ICC staining; imaging DAPI/FITC/CY5
DNA | Albumin | BSEP DNA | ALB | BSEP ICC; DNA (DAPI), Albumin (FITC), BSEP (CY5)
DNA | Albumin | VE-Cadherin DNA|ALB|VEC ICC/IHC imaging targets
DNA | Albumin | ZO-1 DNA|ALB|ZO1 Imaging stain target
DNA | BSEP | CD68 DNA | BSEP | CD68 ICC; DNA (DAPI), BSEP (FITC), CD68 (CY5)
DNA | CD68 | BSEP DNA | CD68 | BSEP ICC; DNA: DAPI, CD68: FITC, BSEP: CY5
DNA | CD68 | CYP3A4 DNA | CD68 | CYP3A4 ICC; DNA: DAPI, CD68: FITC, CYP3A4: CY5
DNA | CYP3A4 | ZO-1 DNA|CYP3A4|ZO1 ICC/IHC imaging target
DNA | F-Actin | ABCB11/BSEP DNA | F-Actin | ABCB IHC/ICC imaging
DNA | F-actin | Albumin DNA | F-actin | Alb ICC; DNA (DAPI), F-actin (FITC), Albumin (CY5)
DNA | F-Actin | alpha 1 Antitrypsin DNA | F-Actin | alph IHC/ICC imaging
DNA | F-Actin | CD68 DNA | F-Actin | CD68 ICC/IHC Imaging targets
DNA | F-Actin | E-Cadherin DNA|F-Actin|E-Cad IHC/ICC imaging targets
DNA | F-Actin | Human Albumin DNA | F-Actin | ALB IHC/ICC imaging
DNA | F-Actin | MDR1/ABCB1 DNA | F-Actin | MDR1 IHC/ICC imaging
DNA | F-Actin | MDR3 DNA | F-Actin | MDR3 IHC/ICC imaging
DNA | F-Actin | MRP1 DNA | F-Actin | MRP ICC/IHC imaging
DNA | F-Actin | MRP2 DNA | F-Actin | MRP2 IHC/ICC imaging
DNA | F-Actin | NTCP/SLC10A1 DNA | F-Actin | NTCP IHC/ICC imaging
DNA | F-Actin | OATP1B1 DNA | F-Actin | OAT ICC/IHC imaging targets
DNA | F-Actin | VE-Cadherin DNA | F-Actin | VE-C ICC/IHC Imaging targets
DNA | GFAP | αSMA DNA | GFAP | αSMA ICC; DNA (DAPI), GFAP (FITC), αSMA (CY5)
DNA | OAT1 DNA | OAT1 ICC / IHC imaging
DNA | OAT1 | ZO-1 DNA | OAT1 | ZO-1 Imaging of DNA, OAT1, and ZO-1
DNA | OAT1 | ZO-1 || DNA | CD31 | VE-Cadherin DNA|OAT1|ZO-1|CD31|V IHC / ICC imaging with different stains in different chambers Top (RPTEC): ZO-1 (green), OAT1 (red), DNA (blue) Bottom (HUVEC): CD31 (green), VE-cadherin (Red), DNA (blue)
DNA | OAT3 DNA | OAT3 ICC / IHC Imaging
DNA | OAT3 | ZO-1 DNA | OAT3 | ZO-1 ICC/IHC imaging
DNA | OAT4 | HO-1 Imm combo 3 Immunocytochemistry imaging.
DNA | OAT4 in Cells | HO-1 in Cells | Tissue Structure Imm combo 4 Immunocytochemistry imaging.
DNA | Occludin | CD68 DNA|Occludin|CD68 IHC/ICC imaging
DNA | OCT 2 DNA | OCT 2 ICC / IHC Imaging
DNA | OCT2 | ZO-1 DNA | OCT2 | ZO-1 Imaging of DNA, OCT2, and ZO-1
DNA | OCT2 | ZO-1 || DNA | CD31 | VE-cadherin DNA|OCT2|ZO1|CD31|VE ICC/ IHC zstack of the chip in the mimetas plate. Top (RPTEC): ZO-1 (green), OCT2 (red), DNA (blue) Bottom (HUVEC): CD31 (green), VE-cadherin (Red), DNA (blue)
DNA | Stabilin 2 | CD31 DNA | Stabilin 2 | C ICC; DNA (DAPI), Stabilin 2 (FITC), CD31 (CY5)
DNA | ZO-1 DNA | ZO-1 Imaging of DNA and ZO-1
DNA | ZO-1 | b-tubulin DNA | ZO-1 | b-tubul Staining for DNA, ZO-1 and b-tubulin
DNA | ZO-1 | OAT3 || DNA | CD31 | VE-cadherin DNA|ZO1|OAT3|CD31|VE IHC/ICC zstack of the chip in the mimetas plate. Top (RPTEC): ZO-1 (green), OAT3 (red), DNA (blue) Bottom (HUVEC): CD31 (green), VE-cadherin (Red), DNA (blue)
HO-1 in Cells HO-1 Immunocytochemistry imaging.
Human Alkaline Phosphatase Human ALKP Human Alkaline Phosphatase
Image IMG Image for later analysis of intensity, morphology, distribution, location, etc.
Laminin Laminin Laminins are a major constituent of the basement membrane which is an intricate meshwork of proteins separating the epithelium, mesothelium, and endothelium from connective tissue. There are 15 different laminins, each consisting of a unique combination of three subchains. The combination of chains confers some tissue specificity. [D. Guldager Kring Rasmussen, M.A. Karsdal, in Biochemistry of Collagens, Laminins and Elastin, 2016]
Live and Dead cell morphology and distribution Calcein | draq7 Live cells stained with calcein/AM and draq 7, then imaged by transmission and fluorescence microscopy looking and 2D and 3D tissue morphology and to asses number and distribution of live and dead cells
Live Cells LIVE Number or percent of live cells
Mitochondria Mitochondria Imaging of the mitochondria in live cells.
Nephrin Nephrin Quantification of Nephrin secretion into cell culture/supernatant
Neutral lipids Neutral lipids Neutral lipids observed in an image using LipidTox Green.
Nuclei Nuclei Nuclei as observed by image staining.
Nucleus Nucleus Nucleus of a cell. Typically this target is used when describing an image with a nuclear stain
Nucleus | Nephrin | CD34 N|Nephrin|CD34 Imaging target for the combined staining of nuclei, CD34, and nephrin
OAT1 OAT1 Oat1 [Slc22a6, originally identified as NKT (novel kidney transporter)] plays a central role in the metabolism and handling of gut microbiome metabolites as well as putative uremic toxins of kidney disease. The organic anion transporter (OAT) subfamily has a role in handling of common drugs (antibiotics, antivirals, diuretics, nonsteroidal anti-inflammatory drugs), toxins (mercury, aristolochic acid), and nutrients (vitamins, flavonoids). [Nigam SK, Bush KT, Martovetsky G, et al. The organic anion transporter (OAT) family: a systems biology perspective. Physiol Rev. 2015;95(1):83-123. doi:10.1152/physrev.00025.2013]
OAT4 in Cells OAT4 Immunocytochemistry imaging.
OCT-2 Quantification (Imaging) OCT-2 Image based quantification of OCT-2 staining
Tissue Structure Tissue Structure Tissue structure as observed by image staining.
VE-Cadherin VE-Cadherin VE-cadherin belongs to the super-family of classical cadherins, and as such, mediates homotypic calcium-dependent cell–cell interactions.1 VE-cadherin expression is tissue-specific and exclusive to endothelial cells. Also know as Cadherin 5, type 2, or CD144 (Cluster of Differentiation 144). [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4049861/]
Vimentin Vimentin EMT marker
ZO-1 ZO-1 Recommended name: Tight junction protein ZO-1 Alternative name(s): Tight junction protein 1 Zona occludens protein 1 Zonula occludens protein 1 TJP1, TJP2, and TJP3 are closely related scaffolding proteins that link tight junction (TJ) transmembrane proteins such as claudins, junctional adhesion molecules, and occludin to the actin cytoskeleton (PubMed:7798316, PubMed:9792688). [https://www.uniprot.org/uniprot/Q07157]

Studies Utilizing this Method/Kit
Study Start Date Study Types Description
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