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Neuronal Marker IF Antibody Sampler Kit #8572

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
GFAP (GA5) Mouse mAb #3670	40 µl	W IHC-P IF-F	H M R	50	Mouse IgG1
CNPase (D83E10) XP® Rabbit mAb #5664	40 µl	W IP IF-F	H M R	47	Rabbit IgG
β3-Tubulin (D71G9) XP® Rabbit mAb #5568	40 µl	W IP IF-F IF-IC	H M R	55	Rabbit IgG
Nestin (Rat-401) Mouse mAb #4760	40 µl	IHC-P IF-F	R	207	Mouse IgG1
Neurofilament-L (C28E10) Rabbit mAb #2837	40 µl	W IHC-P IF-F	H M R	70	Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-F=Immunofluorescence (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Neuronal Marker IF Antibody Sampler Kit has been validated for IF-F, recognizes only its specific target, and does not cross-react with other family members. CNPase (D83E10) XP^® Rabbit mAb recognizes endogenous levels of total CNPase protein. GFAP (GA5) Mouse mAb detects endogenous levels of total GFAP protein. Nestin (Rat-401) Mouse mAb detects endogenous levels of nestin 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. This antibody does not cross-react with tubulin isoforms expressed in non-neuronal cells. This clone is similar to TUJ1. Expression of these proteins may vary in different cells and tissues.

Description

The Neuronal Marker IF Antibody Sampler Kit provides an economical means for labeling neuronal structures by immunofluorescence (IF-F). This kit includes enough primary antibody to perform at least forty IF-F tests or four western blot experiments per primary antibody.

Source / Purification

Rabbit monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of human β3-tubulin protein, or residues surrounding Val81 of human CNPase protein, or Glu450 of human Neurofilament-L protein. Mouse monoclonal antibodies are produced by immunizing animals with E15 rat spinal cord or with native GFAP purified from pig spinal cord.

Background

The antibodies in this kit serve as neuronal markers to determine protein localization in neurons. The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments, and microtubules. Neurofilaments are the major intermediate filaments found in neurons and consist of light (NFL), medium (NFM), and heavy (NFH) subunits (1). Nestin is an intermediate filament family member protein that is structurally related to the neurofilament proteins (2). Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells (3). High CNPase expression is seen in oligodendrocytes and Schwann cells as CNPase accounts for roughly 4% of the total myelin protein in the central nervous system (4). CNPase binds to tubulin heterodimers and plays a role in tubulin polymerization and oligodendrocyte process outgrowth (5). GFAP filaments are characteristic of differentiated and mature brain astrocytes. Thus, GFAP is commonly used by investigators as a marker for intracranial and intraspinal tumors arising from astrocytes (6).

1. Al-Chalabi, A. and Miller, C.C. (2003) Bioessays 25, 346-55.
2. Michalczyk, K. and Ziman, M. (2005) Histol Histopathol 20, 665-71.
3. Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.
4. Kozlov, G. et al. (2003) J Biol Chem 278, 46021-8.
5. Lee, J. et al. (2005) J Cell Biol 170, 661-73.
6. Goebel, H.H. et al. (1987) Acta Histochem Suppl 34, 81-93.

Application References

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Protocols

2837:

IHC / Paraffin, Immunofluorescence, Western Blotting

3670:

IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

4760:

IHC / Paraffin, Immunofluorescence

5568:

Immunofluorescence, Immunoprecipitation, Western Blotting

5664:

Immunofluorescence*, Immunoprecipitation, Western Blotting

* Product-specific protocol.

Companion Products

• 4412 Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate)
• 4414 Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 647 Conjugate)
• 4413 Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 555 Conjugate)
• 8889 Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 594 Conjugate)
• 4408 Anti-mouse IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate)
• 8890 Anti-mouse IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 594 Conjugate)
• 4409 Anti-mouse IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 555 Conjugate)
• 4410 Anti-mouse IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 647 Conjugate)
• 4084 DRAQ5^®
• 9071 ProLong^® Gold Antifade Reagent
• 8961 ProLong^® Gold Antifade Reagent with DAPI
• 8112 SignalStain^® Antibody Diluent
• 8114 SignalStain^® Boost IHC Detection Reagent (HRP, Rabbit)
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7076 Anti-mouse IgG, HRP-linked Antibody
• 9998 BSA
• 9999 Nonfat Dry Milk

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patents No. 5,675,063) from Epitomics, Inc. 

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For Research Use Only. Not For Use In Diagnostic Procedures.