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

Mature Neuron Marker Antibody Sampler Kit #35574

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other Image 1 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 2 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 3 - Mature Neuron Marker Antibody Sampler Kit

Confocal immunofluorescent analysis of frozen rat cerebellum using MAP2 (D5G1) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

other Image 4 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 5 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 6 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 7 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 8 - Mature Neuron Marker Antibody Sampler Kit

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.

other Image 9 - Mature Neuron Marker Antibody Sampler Kit

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.

other Image 10 - Mature Neuron Marker Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded mouse cerebellum using NeuN (D4G4O) XP® Rabbit mAb.

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Western blot analysis of extracts from mouse neonatal brain and rat fetal brain using GAP43 (D9C8) Rabbit mAb.

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Western blot analysis of extracts from neonatal and adult mouse brain using MAP2 (D5G1) XP® Rabbit mAb.

other Image 13 - Mature Neuron Marker Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded mouse brain using Neurofilament-L (C28E10) Rabbit mAb.

other Image 14 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 15 - Mature Neuron Marker Antibody Sampler Kit

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).

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Western blot analysis of extracts from human cerebellum and rat brain using PSD95 (D27E11) XP® Rabbit mAb.

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Confocal immunofluorescent analysis of mouse brain using UCHL1 (D3T2E) XP® Rabbit mAb (green). Blue = Hoescht 33342 #4082 (fluorescent DNA dye).

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Immunohistochemical analysis of paraffin-embedded mouse colon (myenteric plexus) using NeuN (D4G4O) XP® Rabbit mAb.

other Image 19 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 20 - Mature Neuron Marker Antibody Sampler Kit

Western blot analysis of HeLa and human cerebellum using β3-Tubulin (D71G9) XP® Rabbit mAb.

other Image 21 - Mature Neuron Marker Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded Neuro-2a cell pellet (left, positive) or C2C12 cell pellet (right, negative) using Synaptophysin (D8F6H) XP® Rabbit mAb.

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Confocal immunofluorescent analysis of mouse brain using UCHL1 (D8R2I) XP® Rabbit mAb (green). Blue = Hoescht 33342 #4082 (fluorescent DNA dye).

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Immunohistochemical analysis of paraffin-embedded human glioblastoma multiform using NeuN (D4G4O) XP® Rabbit mAb.

other Image 24 - Mature Neuron Marker Antibody Sampler Kit

Western blot analysis of extracts from mouse brain, HeLa cells and rat brain, using Neurofilament-L (C28E10) Rabbit mAb.

other Image 25 - Mature Neuron Marker Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded mouse cerebellum using Synaptophysin (D8F6H) XP® Rabbit mAb.

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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).

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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).

other Image 28 - Mature Neuron Marker Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded mouse colon using Synaptophysin (D8F6H) XP® Rabbit mAb.

other Image 29 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 30 - Mature Neuron Marker Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Synaptophysin (D8F6H) XP® Rabbit mAb.

other Image 31 - Mature Neuron Marker Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using UCHL1 (D3T2E) XP® Rabbit mAb.

other Image 32 - Mature Neuron Marker Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Synaptophysin (D8F6H) XP® Rabbit mAb.

other Image 33 - Mature Neuron Marker Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded DU 145 (left; positive) and LNCaP (right; negative) cell pellets using UCHL1 (D3T2E) XP® Rabbit mAb.

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Immunohistochemical analysis of paraffin-embedded mouse pancreas using Synaptophysin (D8F6H) XP® Rabbit mAb.

other Image 35 - Mature Neuron Marker Antibody Sampler Kit

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).

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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).

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Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using UCHL1 (D3T2E) XP® Rabbit mAb.

other Image 38 - Mature Neuron Marker Antibody Sampler Kit

Western blot analysis of extracts from various cell lines using UCHL1 (D3T2E) XP® Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

other Image 39 - Mature Neuron Marker Antibody Sampler Kit

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).

other Image 40 - Mature Neuron Marker Antibody Sampler Kit

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.

To Purchase # 35574T
Product # Size Price
35574T
1 Kit  (8 x 20 µl) $ 554

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. Sanchez, C. et al. (2000) Prog. Neurobiol. 61, 133-168.
  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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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.