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35574
Mature Neuron Marker Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit
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Mature Neuron Marker Antibody Sampler Kit #35574

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Mature Neuron Marker Antibody Sampler Kit: Image 1
Western blot analysis of extracts from various cell lines using UCHL1 (D3T2E) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Mature Neuron Marker Antibody Sampler Kit: Image 2
Western blot analysis of extracts from mouse brain, C2C12 cells, and rat brain using NeuN (D4G4O) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mature Neuron Marker Antibody Sampler Kit: Image 3
Western blot analysis of extracts from mouse brain, HeLa cells and rat brain, using Neurofilament-L (C28E10) Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 4
Western blot analysis of extracts from human cerebellum and rat brain using PSD95 (D27E11) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 5
Western blot analysis of extracts from mouse and rat tissue using Synaptophysin (D8F6H) XP® Rabbit mAb (upper) and B-Actin (D6A8) Rabbit mAb #8457 (lower).
Mature Neuron Marker Antibody Sampler Kit: Image 6
Western blot analysis of HeLa and human cerebellum using β3-Tubulin (D71G9) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 7
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Mature Neuron Marker Antibody Sampler Kit: Image 8
Western blot analysis of extracts from neonatal and adult mouse brain using MAP2 (D5G1) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 9
Western blot analysis of extracts from mouse neonatal brain and rat fetal brain using GAP43 (D9C8) Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 10
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human UCHL1 (hUCHL1-Myc/DDK; +), Myc/DDK-tagged full-length human UCHL3 (hUCHL3-Myc/DDK; +), Myc/DDK-tagged full-length human UCHL5 (hUCHL5-Myc/DDK; +), and Myc/DDK-tagged full-length human BAP1 (hBAP1-Myc/DDK; +), using UCHL1 (D3T2E) XP® Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).
Mature Neuron Marker Antibody Sampler Kit: Image 11
Immunohistochemical analysis of paraffin-embedded mouse cerebellum using NeuN (D4G4O) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 12
Immunohistochemical analysis of paraffin-embedded mouse brain using Neurofilament-L (C28E10) Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 13
Confocal immunofluorescent analysis of rat cerebellum and retina using PSD95 (D27E11) XP® Rabbit mAb (red) and Neurofilament-L (DA2) Mouse mAb #2835 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 14
Immunohistochemical analysis of paraffin-embedded Neuro-2a cell pellet (left, positive) or C2C12 cell pellet (right, negative) using Synaptophysin (D8F6H) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 15
Confocal immunofluorescent analysis of mouse cerebellum using β3-Tubulin (D71G9) XP® Rabbit mAb (green) and Tau (Tau46) Mouse mAb #4019 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 16
Confocal immunofluorescent analysis of frozen rat cerebellum using MAP2 (D5G1) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 17
Confocal immunofluorescent analysis of rat hippocampus (left) and retina (right) using GAP43 (D9C8) Rabbit mAb (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 18
Western blot analysis of extracts from Ramos and K-562 cells using UCHL1 (D3T2E) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). As expected, UCHL1 protein expression is not detected in K-562 cells.
Mature Neuron Marker Antibody Sampler Kit: Image 19
Immunohistochemical analysis of paraffin-embedded mouse colon (myenteric plexus) using NeuN (D4G4O) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded human brain using Neurofilament-L (C28E10) Rabbit mAb in the presence of control peptide (left) or Neurofilament-L blocking peptide #1005 (right).
Mature Neuron Marker Antibody Sampler Kit: Image 21
Immunohistochemical analysis of paraffin-embedded mouse cerebellum using Synaptophysin (D8F6H) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 22
Confocal immunofluorescent analysis of P19 cells that were differentiated with retinoic acid, using β3-Tubulin (D71G9) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5 #4084® (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 23
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using UCHL1 (D3T2E) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 24
Immunohistochemical analysis of paraffin-embedded human glioblastoma multiform using NeuN (D4G4O) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 25
Confocal immunofluorescent analysis of normal rat cerebellum using Neurofilament-L (C28E10) Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 26
Immunohistochemical analysis of paraffin-embedded mouse colon using Synaptophysin (D8F6H) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 27
Immunohistochemical analysis of paraffin-embedded DU 145 (left; positive) and LNCaP (right; negative) cell pellets using UCHL1 (D3T2E) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 28
Confocal immunofluorescent analysis of mouse hippocampus (left), cortex (middle), and cerebellum (right) using NeuN (D4G4O) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 29
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Synaptophysin (D8F6H) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 30
Immunoprecipitation of β3-Tubulin from mouse brain tissue extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is β3-Tubulin (D71G9) XP® Rabbit mAb. Western blot analysis was performed using β3-Tubulin (D71G9) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as the secondary antibody.
Mature Neuron Marker Antibody Sampler Kit: Image 31
Immunohistochemical analysis of paraffin-embedded colorectal adenocarcinoma using UCHL1 (D3T2E) XP® Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
Mature Neuron Marker Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Synaptophysin (D8F6H) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 33
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using UCHL1 (D3T2E) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 34
Immunohistochemical analysis of paraffin-embedded mouse pancreas using Synaptophysin (D8F6H) XP® Rabbit mAb.
Mature Neuron Marker Antibody Sampler Kit: Image 35
Confocal immunofluorescent analysis of mouse brain using UCHL1 (D3T2E) XP® Rabbit mAb (green). Blue = Hoescht 33342 #4082 (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 36
Confocal immunofluorescent analysis of mouse retina using Synaptophysin (D8F6H) XP® Rabbit mAb (green), Neurofilament-L (DA2) Mouse mAb #2835 (yellow) and Lamin A/C (4C11) Mouse mAb #4777 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Mature Neuron Marker Antibody Sampler Kit: Image 37
Confocal immunofluorescent analysis of mouse brain using UCHL1 (D8R2I) XP® Rabbit mAb (green). Blue = Hoescht 33342 #4082 (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 38
Confocal immunofluorescent analysis of mouse brain using Synaptophysin (D8F6H) XP® Rabbit mAb (green), Neurofilament-L (DA2) Mouse mAb #2835 (yellow) and Lamin A/C (4C11) Mouse #4777 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Mature Neuron Marker Antibody Sampler Kit: Image 39
Confocal immunofluorescent analysis of mouse olfactory bulb (left) and pons (right) using UCHL1 (D3T2E) XP® Rabbit mAb #13179 (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Mature Neuron Marker Antibody Sampler Kit: Image 40
Confocal immunofluorescent analysis of DU 145 (positive; left) and LNCaP (negative; right) cells using UCHL1 (D3T2E) XP® Rabbit mAb (green). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).
Mature Neuron Marker Antibody Sampler Kit: Image 41
Flow cytometric analysis of K-562 cells (blue) and Ramos cells (green) using UCHL1 (D3T2E) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
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To Purchase # 35574
Cat. # Size Qty. Price
35574T		 1 Kit  (8 x 20 microliters)
$ 592

Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
NeuN (D4G4O) XP® Rabbit mAb 24307 20 µl
  • WB
  • IHC
  • IF
 H M R 46-55 Rabbit IgG
GAP43 (D9C8) Rabbit mAb 8945 20 µl
  • WB
  • IP
  • IF
 H M R 38, 43 Rabbit IgG
MAP2 (D5G1) XP® Rabbit mAb 8707 20 µl
  • WB
  • IP
  • IF
 M R 75, 82, 280 Rabbit IgG
Neurofilament-L (C28E10) Rabbit mAb 2837 20 µl
  • WB
  • IHC
  • IF
 H M R 70 Rabbit IgG
β3-Tubulin (D71G9) XP® Rabbit mAb 5568 20 µl
  • WB
  • IP
  • IF
 H M R 55 Rabbit IgG
Synaptophysin (D8F6H) XP® Rabbit mAb 36406 20 µl
  • WB
  • IHC
  • IF
 H M R 38 Rabbit IgG
PSD95 (D27E11) XP® Rabbit mAb 3450 20 µl
  • WB
  • IF
 H M R 95 Rabbit IgG
UCHL1 (D3T2E) XP® Rabbit mAb 13179 20 µl
  • WB
  • IHC
  • IF
  • F
 H M R Mk 27 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
 Goat 

Product Description

The Mature Neuron Marker Antibody Sampler Kit provides an economical means for detecting mature neuron proteins by western and labeling mature neuronal structures by immunofluorescence (IF). This kit includes enough primary antibodies to perform two western blot experiments or at least forty IF tests per primary antibody.

Specificity / Sensitivity

Each antibody in the Mature Neuron Marker Antibody Sampler Kit has been validated for western and IF, recognizes only its specific target, and does not cross-react with other family members. NeuN (D4G4O) XP® Rabbit mAb recognizes endogenous levels of total NeuN protein. GAP43 (D9C8) Rabbit mAb recognizes endogenous levels of total GAP43 protein. MAP2 (D5G1) XP® Rabbit mAb recognizes endogenous levels of total MAP2 protein. Neurofilament-L (C28E10) Rabbit mAb detects endogenous levels of total Neurofilament-L protein. β3-Tubulin (D71G9) XP® Rabbit mAb detects endogenous levels of total β3-tubulin protein that is expressed throughout neuronal development (5). This antibody does not cross-react with tubulin isoforms expressed in non-neuronal cells. This clone is similar to TUJ1. Synaptophysin (D8F6H) XP® Rabbit mAb recognizes endogenous levels of total Synaptophysin protein. PSD95 (D27E11) XP® Rabbit mAb detects endogenous levels of total PSD95 protein. UCHL1 (D3T2E) XP® Rabbit mAb recognizes endogenous levels of total UCHL1 protein. This antibody does not cross-react with other UCH family members.

Source / Purification

Rabbit monoclonal antibodies are produced by immunizing animals with one of the following antigens: 1) synthetic peptides corresponding to residues surrounding Lys30 of human GAP43 protein, Glu450 of human Neurofilament-L protein, Gly299 of human Synaptophysin protein, or Gln53 of human PSD95 protein, 2) synthetic peptides corresponding to residues near the carboxy terminus of human MAP2 protein, the carboxy terminus of human β3-tubulin protein, or the carboxy terminus of human UCHL1 protein, or 3) recombinant protein specific to the amino terminus of human NeuN protein.

Background

The antibodies in this kit serve to characterize and identify mature neurons. Neural stem cells differentiate into mature post-mitotic neurons that are incapable of cellular division. Several neuron-enriched markers can be used to identify mature neurons. Neuronal nuclei (NeuN, Fox-3, RBFOX3) is a nuclear protein expressed in most post-mitotic neurons of the central and peripheral nervous systems. NeuN is not detected in Purkinje cells, sympathetic ganglion cells, Cajal-Retzius cells, INL retinal cells, inferior olivary, or dentate nucleus neurons (1). This neuronal protein was originally identified by immunoreactivity with a monoclonal antibody also called NeuN. Using MS-analysis, NeuN was later identified as the Fox-3 gene product, which contains an RNA recognition motif and functions as a splicing regulator (2). As neurons mature, they develop elaborate processes like axons and dendrites that are necessary to drive core neuronal functions, including synaptic transmission.

GAP43 is a nervous system specific, growth-associated protein enriched in growth cones and areas of high plasticity (3). GAP43 is integral to growth cone formation, neurite outgrowth, and the development of a functional cerebral cortex (4). The cytoskeleton, which is important in generating neuronal processes, consists of three types of cytosolic fibers: actin microfilaments, intermediate filaments, and microtubules. β3-tubulin is one of six β-tubulin isoforms that make up the building blocks of microtubules (5). Microtubule-associated protein 2 (MAP2) is a neuronal phosphoprotein that regulates the structure and stability of microtubules, neuronal morphogenesis, cytoskeleton dynamics, and organelle trafficking in axons and dendrites (6). MAP2 is preferentially localized to dendrites in cultured neurons (7). Neurofilaments are the major intermediate filaments found in neurons and consist of light (NFL), medium (NFM), and heavy (NFH) subunits (8). Similar in structure to other intermediate filament proteins, neurofilaments have a globular amino-terminal head, a central α-helical rod domain, and a carboxy-terminal tail. A heterotetrameric unit (NFL-NFM and NFL-NFH) forms a protofilament, with eight protofilaments comprising the typical 10 nm intermediate filament (9). Neurofilaments are critical for radial axon growth and determine axon caliber, serving as markers for neuronal axons.

Mature neurons function as cellular mediators of synaptic transmission. Synaptophysin is a neuronal synaptic vesicle glycoprotein (10). Synaptophysin is responsible for targeting synaptobrevin 2/VAMP2 to synaptic vesicles, and is a critical component and marker for the presynaptic fusion complex (11). Postsynaptic Density protein 95 (PSD95) is a member of the membrane-associated guanylate kinase (MAGUK) family of proteins. These family members consist of an amino-terminal variable segment followed by three PDZ domains, an SH3 domain, and an inactive guanylate kinase (GK) domain. PSD95 is a scaffolding protein involved in the assembly and function of mature postsynaptic density complexes (12,13).

Several cellular processes are required to support dynamic functions existing in mature neurons, including protein regulation by protein ubiquitination. Ubiquitin C-terminal hydrolase L1 (UCH-L1) is a deubiquitinating enzyme that is selectively and abundantly expressed in the brain, and its activity is required for normal synaptic function (14).
  1. Mullen, R.J. et al. (1992) Development 116, 201-11.
  2. Kim, K.K. et al. (2009) J Biol Chem 284, 31052-61.
  3. Biewenga, J.E. et al. (1996) Acta Biochim Pol 43, 327-38.
  4. Aigner, L. and Caroni, P. (1993) J Cell Biol 123, 417-29.
  5. Jiang, Y.Q. and Oblinger, M.M. (1992) J Cell Sci 103 (Pt 3), 643-51.
  6. Sánchez, C. et al. (2000) Prog Neurobiol 61, 133-68.
  7. Caceres, A. et al. (1984) Brain Res 315, 314-8.
  8. Al-Chalabi, A. and Miller, C.C. (2003) Bioessays 25, 346-55.
  9. Cohlberg, J.A. et al. (1995) J Biol Chem 270, 9334-9.
  10. Wiedenmann, B. and Franke, W.W. (1985) Cell 41, 1017-28.
  11. Bonanomi, D. et al. (2007) Biochem J 404, 525-34.
  12. Cao, J. et al. (2005) J Cell Biol 168, 117-26.
  13. Chetkovich, D.M. et al. (2002) J Neurosci 22, 6415-25.
  14. Gong, B. et al. (2006) Cell 126, 775-88.

Pathways & Proteins

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.