Revision 4

#48510Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Tyrosine Hydroxylase (E2L6M) Rabbit mAb 58844 20 µl 55-60 kDa Rabbit IgG
GAD2 (D5G2) XP® Rabbit mAb 5843 20 µl 60 kDa Rabbit IgG
VGLUT2 (D7D2H) Rabbit mAb 71555 20 µl 65-70 kDa Rabbit IgG
VGLUT1 (E8L5B) Rabbit mAb 47181 20 µl 62 kDa Rabbit IgG
ChAT (E4F9G) Rabbit mAb 27269 20 µl 71 kDa Rabbit IgG
GAD1 (D1F2M) Rabbit mAb 41318 20 µl 67 kDa Rabbit IgG
5-HTR4 (D8O5K) Rabbit mAb 13690 20 µl 40-140 kDa Rabbit IgG
β2-Adrenergic Receptor (D6H2) Rabbit mAb 8513 20 µl 50-100 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Functional Neuron Marker Antibody Sampler Kit provides an economical means of detecting markers that facilitate phenotyping neurons by function. The kit includes enough antibodies to perform at least two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

Tyrosine hydroxylase (TH) catalyzes the rate-limiting step in the synthesis of the neurotransmitter dopamine and other catecholamines, and acts as a marker for dopaminergic neurons (1,2). Choline O-acetyltransferase (ChAT) is the enzyme that catalyzes synthesis of acetylcholine (ACh) in the central and peripheral nervous system. ChAT is found in high levels within cholinergic neurons and can be used to measure their functional states (3). Vesicular glutamate transporters 1 and 2 (VGLUT1 and VGLUT2) are responsible for transporting the excitatory neurotransmitter glutamate into synaptic vesicles of glutamatergic neurons. VGLUT1 and VGLUT2 are complimentarily expressed and act as markers for glutamatergic neurons (4). Glutamate decarboxylase (GAD) is the main enzyme that synthesizes GABA from glutamate. GABA producing neurons, called GABAergic neurons, utilize GABA as their major inhibitory neurotransmitter with both isoforms of GAD, GAD1, and GAD2, acting as functional markers for these neurons (5). β2-adrenergic receptor (β2AR) is a G protein-coupled receptor (GPCR) that mediates the actions of catecholamines, mainly through stimulation by epinephrine (adrenaline), in the central and peripheral nervous system (6,7). Serotonin receptor 4 (5-HTR4) is an excitatory GPCR that activates the cyclic AMP (cAMP)-PKA pathway (8,9). 5-HTR4 is located post-synaptically on serotonergic neurons (10).

  1. Kumer, S.C. and Vrana, K.E. (1996) J Neurochem 67, 443-62.
  2. Weihe, E. et al. Cell Mol Neurobiol 26, 659-78.
  3. Oda, Y. (1999) Pathol Int 49, 921-37.
  4. Ziegler, D.R. et al. (2002) J Comp Neurol 448, 217-29.
  5. Le, T.N. et al. (2017) J Neurosci 37, 8816-8829.
  6. Wu, Y. et al. (2021) Biomolecules 11, 936. doi: 10.3390/biom11070936.
  7. Wallukat, G. (2002) Herz 27, 683-90.
  8. Nichols, D.E. and Nichols, C.D. (2008) Chem Rev 108, 1614-41.
  9. Karayol, R. et al. (2021) Mol Psychiatry 26, 2334-2349.
  10. Samuels, B.A. et al. (2016) Neuroscientist 22, 26-45.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    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.

    Revision 4
    #48510

    Functional Neuron Marker Antibody Sampler Kit

    Functional Neuron Marker Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human 5-HTR4 (h5-HTR4-Myc/DDK; +), using 5-HTR4 (D8O5K) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from human spinal cord, mouse spinal cord, and mouse kidney using ChAT (E4F9G) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Functional Neuron Marker Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from mouse and rat tissue using GAD1 (D1F2M) Rabbit mAb (upper) and B-Actin (D6A8) Rabbit mAb #8457 (lower).
    Functional Neuron Marker Antibody Sampler Kit: Image 4 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse cerebellum using VGLUT1 (E8L5B) Rabbit mAb (left, green), β3-Tubulin (E9F3E) Mouse mAb #45058 (right, red), and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
    Functional Neuron Marker Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from various tissues using VGLUT1 (E8L5B) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of VGLUT1 protein in rat lung tissue is consistent with the predicted expression pattern.
    Functional Neuron Marker Antibody Sampler Kit: Image 6 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse olfactory bulb using VGLUT1 (E8L5B) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from neonatal mouse and fetal rat brain using GAD2 (D5G2) XP® Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from various tissues and cell lines using Tyrosine Hydroxylase (E2L6M) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Functional Neuron Marker Antibody Sampler Kit: Image 9 Expand Image
    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.
    Functional Neuron Marker Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from mouse brain, mouse lung, rat brain, and human cortex using VGLUT2 (D7D2H) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Functional Neuron Marker Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of ACHN cells or 293T cells, either mock transfected (-) or transfected (+) with a cDNA expression construct encoding human β2-adrenergic receptor (hβ2AR), using β2-Adrenergic Receptor (D6H2) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from NIH/3T3 and PC-12 cells using 5-HTR4 (D8O5K) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of GAD1 protein 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 GAD1 (D1F2M) Rabbit mAb. Western blot analysis was performed using GAD1 (D1F2M) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse hippocampus using VGLUT1 (E8L5B) Rabbit mAb (left, green), β3-Tubulin (E9F3E) Mouse mAb #45058 (right, red), and ProLong Gold Antifade Reagent with DAPI #8961 (right, blue).
    Functional Neuron Marker Antibody Sampler Kit: Image 15 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse cortex using VGLUT1 (E8L5B) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 16 Expand Image
    Confocal immunofluorescent analysis of mouse brain using GAD2 (D5G2) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA marker).
    Functional Neuron Marker Antibody Sampler Kit: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using Tyrosine Hydroxylase (E2L6M) Rabbit mAb (left) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    Functional Neuron Marker Antibody Sampler Kit: Image 18 Expand Image
    Immunoprecipitation of VGLUT2 from mouse brain extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is VGLUT2 (D7D2H) Rabbit mAb. Western blot analysis was performed using VGLUT2 (D7D2H) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 19 Expand Image
    Confocal immunofluorescent analysis of rat retina (left) and cerebellum (right) using GAD1 (D1F2M) Rabbit mAb (green) and GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Functional Neuron Marker Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse eye using VGLUT1 (E8L5B) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded PC-12 cell pellet (left, positive) or KNRK cell pellet (right, negative) using Tyrosine Hydroxylase (E2L6M) Rabbit mAb
    Functional Neuron Marker Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of various paraffin-embedded normal human tissues: brain (left-top), tonsil (middle-top), small intestine (right-top), skin (left-bottom), testis (middle-bottom), and spleen (right-bottom) using VGLUT1 (E8L5B) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Tyrosine Hydroxylase (E2L6M) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human brain (left), mouse cerebellum (middle), or mouse dentate gyrus (right) using VGLUT1 (E8L5B) Rabbit mAb (top) or VGLUT1 Antibody (bottom). These two antibodies detect unique, non-overlapping epitopes on VGLUT1 protein. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining. 
    Functional Neuron Marker Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded rat small intestine using Tyrosine Hydroxylase (E2L6M) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse cerebellum using VGLUT1 (E8L5B) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    Functional Neuron Marker Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using Tyrosine Hydroxylase (E2L6M) Rabbit mAb performed on the Leica BOND Rx.
    Functional Neuron Marker Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded 293T cell pellet, untransfected (left) or mouse VGLUT1 transfected (right), using VGLUT1 (E8L5B) Rabbit mAb.
    Functional Neuron Marker Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse colon using Tyrosine Hydroxylase (E2L6M) Rabbit mAb performed on the Leica BOND Rx.
    Functional Neuron Marker Antibody Sampler Kit: Image 30 Expand Image
    Confocal immunofluorescent analysis of rat striatum using Tyrosine Hydroxylase (E2L6M) Rabbit mAb (green) and Neurofilament-L (DA2) Mouse mAb #2835 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Functional Neuron Marker Antibody Sampler Kit: Image 31 Expand Image
    Confocal immunofluorescent analysis of HCC1419 cells (left, positive) and ACHN cells (right, negative) using Tyrosine Hydroxylase (E2L6M) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue = Hoechst 33342 #4082 (fluorescent DNA dye).
    Functional Neuron Marker Antibody Sampler Kit: Image 32 Expand Image
    Flow cytometric analysis of KNRK cells (blue, negative) and PC-12 cells (green, positive) using Tyrosine Hydroxylase (E2L6M) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.