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Product details

Synonyms = Neural cell adhesion molecule L1, CAML1, HSAS1, Hyd, L1 Cell Adhesion Molecule, L1-NCAM, MASA, MIC5, NCAM-L1, Nerve-growth factor-inducible large external glycoprotein, Neural cell adhesion molecule L1, NILE, S10, SPG1

Antibody type = Recombinant Rabbit monoclonal / IgG

Clone = MSVA-171R

Positive controlKidney: A strong membranous L1CAM staining should be seen in a subset of collecting ducts while the vast majority of tubuli remains L1CAM negative.

Negative controlKidney: L1CAM staining must be absent in the vast majority of tubuli (L1CAM staining is limited to collecting ducts).

Cellular localization = Nucleus

Reactivity = Human

 

 

Application = Immunohistochemistry
Dilution = 1:100 – 1:200
Intended Use = Research Use Only

Relevance of Antibody

CD171 is a critical protein for brain development and cancer progression.

Biology Behind

The cell adhesion molecule L1 (CD171; L1CAM) is a multidomain membrane Typ 1 glycoprotein of the immunoglobulin superfamily coded by the L1CAM gene at chromosome Xq28. It plays a role in the development of the nervous system, by regulation of cell migration, adhesion, neuronal differentiation, myelination, and axon growth. L1 protein is located all over the nervous system on the surface of neurons. It is placed along the cellular membrane so that one end of the protein remains inside the nerve cell while the other end stays on the outer surface of the neuron. This position allows the protein to activate chemical signals which spread through the neuron. L1CAM has also been proposed as a marker of Extracellular Vesicles (EVs) originating from neuronal cells. Mutations in the L1 protein are the cause of L1 syndrome, characterized by corpus callosum hypoplasia, retardation, aphasia, spastic paraplegia and hydrocephalus). L1CAM protein expression also occurs in various types of tumors where it has been associated with tumor progression, metastasis, therapy resistance, stemness, and dismal outcome. L1CAM protein is currently evaluated as a therapeutic target by neutralizing antibodies, radioimmunoconjugates or chimeric antigen receptor-redirected T (CAR-T) cells.

Staining Pattern in Normal Tissues

L1CAM expression predominantly occurs in nerve fibres but it is also seen in few epithelial tissue types.  Images describing the CD171 staining pattern in normal tissues obtained by the antibody MSVA-171R are shown in our “Normal Tissue Gallery”.

Brain Cerebrum Intense L1CAM staining, especially of nerve fibres in the grey matter.
Cerebellum Intense L1CAM staining, especially of nerve fibres in the molecular layer.
Endocrine Tissues Thyroid Negative.
Parathyroid Negative.
Adrenal gland Strong, predominantly membranous L1CAM staining of a medullary cells.
Pituitary gland Intense L1CAM staining of fibres in the neurohypophysis. Moderate L1CAM staining of a subset of epithelial cells in the adenohypophysis.
Respiratory system Respiratory epithelium Negative.
Lung Negative.
Gastrointestinal Tract Salivary glands Negative.
Esophagus Negative.
Stomach Distinct L1CAM staining of nerve fibres in the lamina mucosae.
Duodenum Distinct L1CAM staining of nerve fibres in the lamina mucosae.
Small intestine Distinct L1CAM staining of nerve fibres in the lamina mucosae.
Appendix Distinct L1CAM staining of nerve fibres in the lamina mucosae. Strong L1CAM staining of nerve fibres and ganglia in the muscular wall.
Colon Distinct L1CAM staining of nerve fibres in the lamina mucosae. Strong L1CAM staining of nerve fibres and ganglia in the muscular wall.
Rectum Distinct L1CAM staining of nerve fibres in the lamina mucosae. Strong L1CAM staining of nerve fibres and ganglia in the muscular wall.
Liver Negative.
Gallbladder Negative.
Pancreas Negative.
Genitourinary Kidney Distinct membranous L1CAM staining of collecting duct (and perhaps distal tubulus) cells.
Urothelium Negative.
Male genital Prostate Distinct L1CAM staining of nerve fibres.
Seminal vesicles Distinct L1CAM staining of nerve fibres.
Testis Negative.
Epididymis Negative.
Female genital Breast Negative.
Uterus, myometrium Negative.
Uterus, ectocervix Negative.
Uterus endocervix Negative.
Uterus, endometrium Negative.
Fallopian Tube Strong membranous L1CAM staining of a variable subset of epithelial cells.
Ovary Negative.
Placenta early Negative.
Placenta mature Negative.
Amnion Negative.
Chorion Weak to moderate L1CAM staining.
Skin Epidermis Negative.
Sebaceous glands Negative.
Muscle/connective tissue Heart muscle Strong L1CAM staining of small nerve fibres.
Skeletal muscle Negative.
Smooth muscle Negative.
Vessel walls Negative.
Fat Negative.
Stroma Negative.
Endothelium L1CAM staining of endothelial cells can occasionally be seen.
Bone marrow/lymphoid tissue Bone marrow Negative.
Lymph node Weak to moderate L1CAM staining of subsets of B-cells and monocytic cells, mainly of the germinal centre.
Spleen Negative.
Thymus Negative.
Tonsil Weak to moderate L1CAM staining of subsets of B-cells and monocytic cells, mainly of the germinal centre.
Remarks Distinct L1CAM staining of nerve fibres and ganglia can occur in many tissues

 

MCM2 is a ubiquitously expressed protein. The findings described above are thus consistent with the RNA data described in the Human Protein Atlas (Tissue expression CD171). L1CAM expression predominantly occurs in nerve fibres but it is also seen in few epithelial tissue types.

 

Positive controlKidney: A strong membranous L1CAM staining should be seen in a subset of collecting ducts while the vast majority of tubuli remains L1CAM negative.

Negative controlKidney: L1CAM staining must be absent in the vast majority of tubuli (L1CAM staining is limited to collecting ducts).

 

Normal tissue gallery

Staining Pattern in Relevant Tumor Types

L1CAM is primarily expressed in melanoma, ovarian cancer, endometrial carcinoma, renal cell carcinoma, and urothelial carcinomas but it can also be expressed at significant levels in carcinomas of various other organs of origin.

The TCGA findings on CD171 RNA expression in different tumor categories have been summarized in the Human Protein Atlas.

 

 

Cancer tissue gallery

Compatibility of Antibodies

No data available at the moment

Protocol Recommendations

IHC users have different preferences on how the stains should look like. Some prefer high staining intensity of the target stain and even accept some background. Others favor absolute specificity and lighter target stains. Factors that invariably lead to more intense staining include higher concentration of the antibody and visualization tools, longer incubation time, higher temperature during incubation, higher temperature and longer duration of the heat induced epitope retrieval (slide pretreatment). The impact of the pH during slide pretreatment has variable effects and depends on the antibody and the target protein.

 

All images and data shown here and in our image galleries are obtained by the manual protocol described below. Other protocols resulting in equivalent staining are described as well.

 

Manual protocol

Freshly cut sections should be used (less than 10 days between cutting and staining). Heat-induced antigen retrieval for 5 minutes in an autoclave at 121°C in pH 7,8 Target Retrieval Solution buffer. Apply MSVA-171R at a dilution of 1:150 at 37°C for 60 minutes. Visualization of bound antibody by the EnVision Kit (Dako, Agilent) according to the manufacturer’s directions.

Potential Research Applications

  • The clinical utility of a potential prognostic role of L1CAM expression needs to be further evaluated.
  • The utility of L1CAM is a potential therapeutic target that needs to be further investigated.
  • The diagnostic role of L1CAM IHC is not clear.

Evidence for Antibody Specificity in IHC

There are two ways how the specificity of antibodies can be documented for immunohistochemistry on formalin fixed tissues. These are: 1. Comparison with a second independent method for target expression measurement across a large number of different tissue types (orthogonal strategy), and 2. Comparison with one or several independent antibodies for the same target and showing that all positive staining results are also seen with other antibodies for the same target (independent antibody strategy). 

 

Orthogonal validation: For the antibody MSVA-171R specificity is suggested by the strong concordance of the immunostaining data with data from three independent RNA screening studies, including the Human Protein Atlas (HPA) RNA-seq tissue dataset, the FANTOM5 project, and the Genotype-Tissue Expression (GTEx) project, which are all summarized in the Human Protein Atlas (Tissue expression CD171). L1CAM positivity by MSVA-171R is detectable in all tissues with documented L1CAM RNA expression (adrenal gland, pituitary gland, prostate, kidney, seminal vesicles, gastrointestinal tract). The only tissue without previously documented significant L1CAM RNA expression but L1CAM positivity by MSVA-171R (fallopian tube) had only very few positive cells that were probably not detected in RNA studies analyzing entire organs.

 

Comparison of antibodies: True expression of L1CAM in all cell types found L1CAM positive by MSVA-171R is further corroborated by identical stainings obtained by another commercially available independent antibody (termed “validation antibody”). This validation also confirmed positivity of a subset of epithelial cells in the fallopian tube.



 

 

Normal tissue gallery