Prestige Monoclonals as Neuroscience Markers

The Neuroscience Marker Panel

The nervous system is the system that controls and regulates both voluntary and involuntary processes, and higher-order functions, such as perception, cognition, emotions and others.

The principal cells of the nervous system are the neurons, which process and transmit neural signals. In addition, different types of glial cells are present, including astrocytes, oligodendrocytes and Schwann cells that support, nourish and provide electrical isolation to the neural processes. The complexity of the nervous system is further augmented by the fact that different types of neurons utilize various types of chemical substances to transmit the information (chemical neurotransmitters), and that multiple receptor subtypes are present.

Antibody-based immunohistochemistry is widely used in neuroscience research to detect and characterize various cell types in the nervous system, and the need for highly-characterized and specific antibodies is still high.

Sigma-Aldrich has partnered with Atlas Antibodies, who developed a novel panel of Prestige Monoclonal antibodies designed to recognize the main anatomical and neurochemical cell types in rodent and human nervous system. We have taken a great care to be able to offer these markers as tools for mapping the structures and cell types in the central and peripheral nervous system:

  • •Selected target proteins are expressed only by a single cell type
  • IHC-validation in rat, mouse and human tissues
  • WB-validation in mouse and human tissue lysates for the majority of the markers
  • Antibodies are available with different isotypes, allowing for multiplexing experiments
  • Information on epitope and antigen used for immunization is provided (when available)

The monoclonal antibodies within the panel have been developed under the same stringent conditions as all Monoclonal Prestige Antibodies® guaranteeing a secured continuity and stable supply.

Markers for Neural Lineage and Signaling

The Neuroscience Marker panel consists of neural lineage markers for neurons, astrocytes and oligodendrocytes/Schwann cells. The signaling markers target glutamate, GABA, acetylcholine, noradrenaline, dopamine and serotonin systems.

The three different cell types, including neurons, oligodendrocytes and astrocytes, are shown in Figure 2. The antibodies used are Anti-NEFM (AMAb91030), Anti-CNP (AMAb91068) and Anti-GFAP (AMAb91033) detected with isotype-specific secondary antibodies.

Some of the major brain neurotransmitter systems are shown in Figure 3. The image demonstrates the GABAergic, glutamatergic and acetylcholine systems, here visualized by Anti-GAD1 (AMAb91076), Anti-VGLUT1 (AMAb91041) and Anti-CHAT (AMAb91129) antibodies, respectively. Isotype-specific secondary antibodies were used for detection.

High Specificity and Interspecies Reactivity

Prestige Monoclonals Neuroscience markers show high specificity and selectivity for their target proteins. In Figure 4, there is an example of the Anti-NET (AMAb91116) monoclonal antibody. This antibody recognizes the noradrenaline transporter (NET, SLC6A2) and can be used to detect both noradrenergic cell bodies and processes in rat, mouse and human nervous system. The Anti-NET antibody AMAb91116 is highly-specific and does not show any cross-reactivity with e.g. dopamine transporter (SLC6A3, DAT).

Figure 4 shows specific staining of noradrenergic cell bodies and fibers in rat locus coeruleus (A), noradrenergic fibers in mouse cerebral cortex (B), and noradrenergic cell bodies and fibers in human locus coeruleus (C). The specificity of the AMAb91116 is further demonstrated on images D and E. These images show a coronal section of rat brain at the level of caudate putamen stained with Anti-NET (AMAb91116) and Anti-DAT (AMAb91125). The caudate putamen is virtually devoid of noradrenaline fibers, which are mostly present in cortex. A dense network of thin dopamine fibers (in magenta) is present in striatum, where single noradrenaline fibers can be observed sometimes. (in green).

Multiplexed IHC-IF staining of a coronal section of rat brain visualizing neurons in green, oligodendrocytes in magenta and astrocytes in red.

Figure 1. Multiplexed IHC-IF staining of a coronal section of rat brain visualizing neurons in green, oligodendrocytes in magenta and astrocytes in red. Anti-NEFM antibody of isotype IgG2b (AMAb91030) is used to show neurons and their processes, oligodendrocytes are detected by Anti-CNP antibody of isotype IgG2a (AMAb91068) and astrocytes by Anti-GFAP antibody of isotype IgG1 (AMAb91033).
 

Multiplexed IHC-IF staining of mouse caudate putamen/globus pallidus section showing the GABAergic system in red, glutamatergic system in green and acetylcholine system in magenta.

Figure 2. Multiplexed IHC-IF staining of mouse caudate putamen/globus pallidus section showing the GABAergic system in red, glutamatergic system in green and acetylcholine system in magenta. Anti-GAD1 antibody of isotype IgG2a (AMAb91076) is used as marker for the GABAergic system, Anti-VGLUT1 antibody of isotype IgG2b (AMAb91041) for the glutamatergic system and Anti-CHAT antibody of isotype IgG1 (AMAb91129) is used to visualize the acetylcholine system.
 

IHC-IF (A, B, D, E) and bright-field IHC (C) stains demonstrating specificity and selectivity of Anti-NET antibody (AMAb91116) in rat (A, D, E), mouse (B) and human (C) brain.

Figure 3. IHC-IF (A, B, D, E) and bright-field IHC (C) stains demonstrating specificity and selectivity of Anti-NET antibody (AMAb91116) in rat (A, D, E), mouse (B) and human (C) brain. Staining with Anti-NET (AMAb91116) is shown in green (A, B, D, E) and in brown (C). DAT immunoreactivity is visualized in magenta using Anti-DAT antibody (AMAb91125).
 

Table 1.
Summary of the Prestige Monoclonals Neuroscience Markers

 

Marker for Product Name Product No. Validated
Applications
Epitope Sequence identity
mouse/rat
Isotype
Neurons Anti-NEFM (NF160) AMAb91027 IHC*, WB* SDHLEEDIHRLKERF 98/98% IgG1 K
Neurons Anti-NEFM (NF160) AMAb91028 IHC*, WB* LKERFEEEARLRDDT 98/98% IgG1 K
Neurons Anti-NEFM (NF160) AMAb91029 IHC*, WB* SDHLEEDIHRLKERF 98/98% IgG2a K
Neurons Anti-NEFM (NF160) AMAb91030 IHC*, WB* SDHLEEDIHRLKERF 98/98% IgG2b K
Neurons Anti-NEFH (NF200) AMAb91025 IHC, WB ECRIGFGPIPFSLPE 88/94% IgG1 K
Neurons Anti-UCHL1 (PGP9.5) AMAb91145 IHC*, WB* AVANNQDKLG 97/97% IgG1
Astrocytes Anti-GFAP AMAb91033 IHC*, WB* PVQTFSNLQIRETSL 98/100% IgG1 K
Astrocytes Anti-S100B AMAb91038 IHC*, WB N.D. 99/98% IgG1 K
Astrocytes Anti-GLUL AMAb91101 IHC*, WB* LCEVFKYNRR 95/92% IgG1
Astrocytes Anti-GLUL AMAb91102 IHC*, WB* AMFRDPFRKD 95/92% IgG1
Astrocytes Anti-GLUL AMAb91103 IHC*, WB* YLVPAAMFRDPFRKD 95/92% IgG2a K
Schwann cells, oligodendrocytes Anti-MBP AMAb91062 IHC*, WB* RTPPPSQGKG 97/97% IgG2a K
Schwann cells, oligodendrocytes Anti-MBP AMAb91063 IHC*, WB* N.D. 97/97% IgG1
Schwann cells, oligodendrocytes Anti-MBP AMAb91064 IHC*, WB* RTPPPSQGKG 97/97% IgG1
Oligodendrocytes Anti-MOG AMAb91066 IHC*, WB FSRVVHLYRNGKDQD 91/89% IgG1
Oligodendrocytes Anti-MOG AMAb91067 IHC*, WB GMEVGWYRPP 91/89% IgG1
Oligodendrocytes Anti-CNP AMAb91068 IHC*, WB* PGVLHCTTKFCDYGK 76/77% IgG2a K
Oligodendrocytes Anti-CNP AMAb91069 IHC*, WB* N.D. 76/77% IgG1
Oligodendrocytes Anti-CNP AMAb91072 IHC*, WB* LSEQQLQLWPSDVDK 76/77% IgG2b K
Acetylcholine neurons Anti-CHAT AMAb91130 IHC* GLFSSYRLPGHTQDT 96/96% IgG2b
Acetylcholine neurons Anti-CHAT AMAb91129 IHC* GLFSSYRLPGHTQDT 96/96% IgG1
Glutamate neurons Anti-SLC17A7 (VGLUT1) AMAb91041 IHC*, WB EERKYIEDAI 94/94% IgG2b K
Glutamate neurons Anti-SLC17A6 (VGLUT2) AMAb91081 IHC* EETGDITQNYINYGT 85/85% IgG1
Glutamate neurons Anti-SLC17A6 (VGLUT2) AMAb91086 IHC* NGGWPSGWEK 85/85% IgG1
GABA neurons Anti-SLC32A1 (VGAT ) AMAb91043 IHC* N.D. 95/93% IgG1 λ
GABA neurons Anti-GAD1 (GAD67) AMAb91076 IHC*, WB TETDFSNLFA 94/94% IgG2a K
GABA neurons Anti-GAD1 (GAD67) AMAb91078 IHC*, WB TETDFSNLFA 94/94% IgG1
GABA neurons Anti-GAD1 (GAD67) AMAb91079 IHC*, WB* TETDFSNLFA 94/94% IgG2b K
GABA neurons Anti-GAD2 (GAD65) AMAb91048 IHC*, WB* AACACDQKPC 84/88% IgG1 K
Dopamine neurons Anti-SLC6A3 (DAT) AMAb91125 IHC* ELPWIHCNNSWNSPN 85/85% IgG1
Dopamine neurons Anti-DDC AMAb91089 IHC*, WB MDWLGKMLEL 90/88% IgG1
Noradrenaline neurons Anti-SLC6A2 (NET) AMAb91116 IHC* HTKYSKYKFTPAAEF N.D. IgG1
Dopamine and noradrenaline neurons Anti-TH AMAb91112 IHC* N.D. 88/88% IgG1
Serotonin neurons Anti-TPH2 AMAb91108 IHC* N.D. 100/100% IgG1