Using Antibodies to Detect the Expression of Neural Stem Cell Markers

by Atlas Antibodies

Neural Stem Cells

Collectively, markers against neural stem cells (NSCs) represent a robust tool in basic research and advanced regenerative medicine. Studies focused on differentiation toward specific neural lineages are supported by changes in the expression levels of specific markers, which can help to identify the presence of neural stem cells. Nestin and SOX2 are two commonly utilized markers for NSCs. Other markers expressed at the cellsurface are ABCG2, FGFR1 and Frizzled-9. NSCs differentiate into neuronsoligodendrocytes and astrocytes (see schematic in figure 1)Each class of stem cells has specific markers. Tables 1-4 contain a list of selected NSC markers available from Atlas Antibodies.

Neurons

The neurons (nerve cells) are electrically responsive cells that communicate and transfer the nerve signal via synapses within the central nervous system and in the periphery. All neurons fall into various functional classes according to their neurochemical characteristics. An example of neuronal marker in human and rodent brain is show in figure 2.

Astrocytes

Astrocytes comprise the principal glial cell population in the central nervous system. These cells derive from divergent populations of progenitor cells in the neuroepithelium of the evolving central nervous system. Astrocytes play various functional roles in the brain, such as the secretion or absorption of neural transmitters, ion homeostasis and maintenance of the blood–brain barrier. An example of astroglial marker in human and rodent brain is show in figure 3.

Oligodendrocytes

Oligodendrocytes are glial cells found inside the CNS and spinal cord. The main function of these cells is to constitute the insulating myelin sheaths located among the neuronal axons. Oligodendrocytes produce different growth factors, such as BDNF and GDNF, thus supplying neurons with trophic assistance. An example of oligodendroglial marker in human and rodent brain is show in figure 4.

Figure 1. NSCs primarily differentiate into neurons, astrocytes, and oligodendrocytes, depending on environmental cues.
Representative staining of Neurofilament using monoclonal Anti-NEFL antibody (AMAb91314). (A) The human cerebellum shows strong cytoplasmic immunoreactivity in cell bodies and proximal dendrites of Purkinje cells. (B) Immunofluorescence staining of mouse striatum shows strong positivity in neural fibers. See Table 2. for a list of Neuronal Stem Cell markers available from Atlas Antibodies.

Figure 2. Representative staining of Neurofilament using monoclonal Anti-NEFL antibody (AMAb91314). (A) The human cerebellum shows strong cytoplasmic immunoreactivity in cell bodies and proximal dendrites of Purkinje cells. (B)Immunofluorescence staining of mouse striatum shows strong positivity in neural fibers. See Table 2. for a list of Neuronal Stem Cell markers available from Atlas Antibodies.

Representative staining of Glial Fibrillary Acidic Protein, GFAP, using Anti-GFAP monoclonal antibody (AMAb91033). (A) Immunohistochemical staining of the human cerebral cortex shows strong cytoplasmic positivity in astrocytes. (B) Immunofluorescence staining of rat hippocampus shows strong positivity of the astrocytes. See Table 3. for a list of Astroglial markers available from Atlas Antibodies.

Figure 3. Representative staining of Glial Fibrillary Acidic Protein, GFAP, using Anti-GFAP monoclonal antibody (AMAb91033). (A) Immunohistochemical staining of the human cerebral cortex shows strong cytoplasmic positivity in astrocytes. (B)Immunofluorescence staining of rat hippocampus shows strong positivity of the astrocytes. See Table 3. for a list of Astroglial markers available from Atlas Antibodies.

Representative staining of Myelin Basic Protein using Anti-MBP monoclonal antibody (AMAb91063). (A) Immunohistochemical staining of the human cerebral cortex shows strong immunoreactivity in myelinated fibers. (B) Immunofluorescence staining in the rat cerebral cortex shows strong immunoreactivity in myelinated axons. See Table 4. for a list of Oligodendroglial markers available from Atlas Antibodies.

Figure 4. Representative staining of Myelin Basic Protein using Anti-MBP monoclonal antibody (AMAb91063). (A) Immunohistochemical staining of the human cerebral cortex shows strong immunoreactivity in myelinated fibers. (B) Immunofluorescence staining in the rat cerebral cortex shows strong immunoreactivity in myelinated axons. See Table 4. for a list of Oligodendroglial markers available from Atlas Antibodies.

NSCs in the Adult Mammalian Brain

Neural progenitor stem cells (NSCs), in the adult mammalian brain, play a key role in developing brain plasticity throughout life.1 When adult NSCs were originally identified, it was thought they merely serves as a regenerative source for new neurons. However, cumulative evidence suggests that the main function of endogenous adult NSCs is to furnish the brain with an extra layer of plasticity, via both direct and indirect  mechanism.2

Adult somatic stem cells offer homeostatic support to the maintenance of tissue organization. Regulation of gene expression by transcription factors is one fundamental mechanism that controls adult NSCs.3 Although apparently without synapse, NSCs express functional receptors and can communicate with multiple neurotransmitters.4-5 In the adult mammalian brain, there are two significant neurogenic of endogenous NSCs: the subventricular zone (SVZ) lining the lateral ventricles and the subgranular zone (SGZ) within the dentate gyrus of the hippocampus. 

Table 1. Neural Stem Cell (NSC) Markers available from Atlas Antibodies

TargetCatalog NoClonalityValidated ApplicationSequence Identity
Mouse/Rat
BLBp/FABP7AMAb90595MonoclonalIHC, WB89%/90%
BMIHPA030472PolyclonalIHC, WB*95%/95%
CD133/PROML1HPA004922PolyclonalIHC*57%/60%
EGFR/ERBB1AMAb90819MonoclonalWB90%/91%
EGR1 / INGFI-AHPA029937PolyclonalICC-IF93%/94%
PCGF5HPA038349PolyclonalIHC, ICC-IF95%/93%
REELIN/RELNHPA046512PolyclonalICC-IF90%/94%
SLC1A3/GLASTHPA037468PolyclonalIHC*93%/93%
TLX/NR2E1HPA055642PolyclonalIHC*99%/99%

Table 2. Neuronal Stem Cell Markers available from Atlas Antibodies

TargetCatalog NoClonalityValidated ApplicationSequence Identity
Mouse/Rat
BRN2/POU3F2AMAb91406MonoclonalIHC, WB, ICC-IF100%/100%
BRN2/POU3F2HPA056261PolyclonalICC-IF100%/100%
FGFR1HPA056402PolyclonalIHC, WB94%/96%
FOXA2HPA050505PolyclonalIHC*, ICC-IF96%/82%
MAP2AMAb91375MonoclonalIHC, WB, ICC-IF91%/90%
NEFLAMAb91314MonoclonalIHC, WB, ICC-IF97%/99%
NURR1/NR4A2HPA000543PolyclonalIHC, ICC-IF100%/100%
PAX3HPA063659PolyclonalIHC, ICC-IF92%/92%
PAX3HPA069000PolyclonalICC-IF98%/98%
PAX6AMAb91372MonoclonalIHC, ICC-IF100%/100%
S100BAMAb91038MonoclonalIHC*, WB99%/98%
SOX11HPA000536PolyclonalIHC, WB82%/82%
SOX21AMAb91309MonoclonalIHC, WB
SOX21HPA048337PolyclonalIHC96%/37%
SOX21HPA064084PolyclonalICC-IF96%/37%
SOX4AMAb91378MonoclonalIHC, ICC-IF
SOX4AMAb91380MonoclonalIHC, ICC-IF
TBR2/EOMESHPA065458PolyclonalICC-IF96%/96%
TUBB3AMAb91394MonoclonalIHC, WB, ICC-IF

Table 3. Astroglial Markers available from Atlas Antibodies

TargetCatalog NoClonalityValidated ApplicationSequence Identity
Mouse/Rat
AQP4AMAb90537MonoclonalIHC*, WB93%/92%
AQP4HPA014784PolyclonalIHC*, WB, ICC-IF93%/92%
AQP9HPA074762PolyclonalIHC*55%/55%
CD44HPA005785PolyclonalIHC*, WB*, ICC-IF51%/47%
GFAPAMAb91033MonoclonalIHC*, WB*98%/100%
GFAPHPA056030PolyclonalIHC*, WB, ICC-IF98%/100%
GFAPHPA063513PolyclonalIHC*100%/98%

Table 4. Oligodendroglial Markers available from Atlas Antibodies

TargetCatalog NoClonalityValidated ApplicationSequence Identity
Mouse/Rat
CD140/PDGFRAHPA004947PolyclonalICC-IF
MBPHPA049222PolyclonalIHC, WB97%/97%
OLIG2HPA003254PolyclonalIHC*, WB93%/94%
PROX1HPA000842PolyclonalICC-IF100%/93%
PROX1HPA001030PolyclonalICC-IF100%/99%
SOX10AMAb91297MonoclonalIHC, ICC-IF98%/98%
SOX8HPA058665PolyclonalIHC*, ICC-IF73%/70%

*Products with enhanced validation for indicated application

See the Atlas Antibodies product catalog for additional product information and ordering details. 

Enhanced Validation

In addition to the extensive validation and characterization always performed, Atlas Antibodies conduct application specific enhanced validation. Enhanced validation offers increased security of antibody specificity in a defined context.

The enhanced validation follows the guidelines proposed by the International Working Group for Antibody Validation (IWGAV). Enhanced validation consists of five conceptual pillars for antibody validation, to be used in an application-specific manner: genetic validation, orthogonal validation, validation by independent antibodies, recombinant expression validation and, migration capture MS validation.

References
  1. Kempermann G. and Gage FH. (1999) Sci. Am. 280: 48-53.
  2. Christian KM.et al.( 2014) Annu. Rev. Neurosci. 37: 243-262
  3. Hsieh J. (2012) Genes Dev. 26: 1010-1021
  4. Berg DA. et al. (2013) Development. 140: 2548-2561
  5. Kunze A. et al. (2009) PNAS, USA. 106: 11336-11341


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