Ms X Neuronal Nuclei (NeuN), Biotin Conjugated

Species Reactivity	Key Applications	Host	Format	Antibody Type
M, R, H, Ft, Ch, Sal	ICC, IHC, IH(P), WB	M	Biotin	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

M, R, H, Ft, Ch, Sal

ICC, IHC, IH(P), WB

Biotin

Monoclonal Antibody

Description
Catalogue Number	MAB377B
Brand Family	Chemicon®
Trade Name	Chemicon
Description	Anti-NeuN Antibody, clone A60, biotin conjugated
Alternate Names	Neuron-Specific Nuclear Protein
Background Information	NeuN antibody (NEUronal Nuclei; clone A60) specifically recognizes the DNA-binding, neuron-specific protein NeuN, which is present in most CNS and PNS neuronal cell types of all vertebrates tested. NeuN protein distributions are apparently restricted to neuronal nuclei, perikarya and some proximal neuronal processes in both fetal and adult brain although, some neurons fail to be recognized by NeuN at all ages: INL retinal cells, Cajal-Retzius cells, Purkinje cells, inferior olivary and dentate nucleus neurons, and sympathetic ganglion cells are examples (Mullen et al., 1992; Wolf et al., 1996). Immunohistochemically detectable NeuN protein first appears at developmental timepoints that correspond with the withdrawal of the neuron from the cell cycle and/or with the initiation of terminal differentiation of the neuron (Mullen et al., 1992). Immunoreactivity appears around E9.5 in the mouse neural tube and is extensive throughout the developing nervous system by E12.5. Strong nuclear staining suggests a nuclear regulatory protein function; however, no evidence currently exists as to whether the NeuN protein antigen has a function in the distal cytoplasm or whether it is merely synthesized there before being transported back into the nucleus. No difference between protein isolated from purified nuclei and whole brain extract on immunoblots has been found (Mullen et al., 1992).

Description

Catalogue Number

MAB377B

Brand Family

Chemicon®

Trade Name

Chemicon

Description

Anti-NeuN Antibody, clone A60, biotin conjugated

Alternate Names

Neuron-Specific Nuclear Protein

Background Information

NeuN antibody (NEUronal Nuclei; clone A60) specifically recognizes the DNA-binding, neuron-specific protein NeuN, which is present in most CNS and PNS neuronal cell types of all vertebrates tested. NeuN protein distributions are apparently restricted to neuronal nuclei, perikarya and some proximal neuronal processes in both fetal and adult brain although, some neurons fail to be recognized by NeuN at all ages: INL retinal cells, Cajal-Retzius cells, Purkinje cells, inferior olivary and dentate nucleus neurons, and sympathetic ganglion cells are examples (Mullen et al., 1992; Wolf et al., 1996). Immunohistochemically detectable NeuN protein first appears at developmental timepoints that correspond with the withdrawal of the neuron from the cell cycle and/or with the initiation of terminal differentiation of the neuron (Mullen et al., 1992). Immunoreactivity appears around E9.5 in the mouse neural tube and is extensive throughout the developing nervous system by E12.5. Strong nuclear staining suggests a nuclear regulatory protein function; however, no evidence currently exists as to whether the NeuN protein antigen has a function in the distal cytoplasm or whether it is merely synthesized there before being transported back into the nucleus. No difference between protein isolated from purified nuclei and whole brain extract on immunoblots has been found (Mullen et al., 1992).

Product Information
Format	Biotin
Control	Brain tissue, most neuronal cell types throughout the adult nervous system
Presentation	Purified mouse monoclonal IgG1 in buffer containing 0.01 M PBS pH 7.1, 0.1% sodium azide with 15 mg/mL BSA as a stabilizer.

Product Information

Format

Biotin

Control

Brain tissue, most neuronal cell types throughout the adult nervous system

Presentation

Purified mouse monoclonal IgG1 in buffer containing 0.01 M PBS pH 7.1, 0.1% sodium azide with 15 mg/mL BSA as a stabilizer.

Applications
Application	This Anti-NeuN Antibody, clone A60, biotin conjugated is validated for use in IC, IH, IH(P), WB for the detection of NeuN.
Key Applications	Immunocytochemistry Immunohistochemistry Immunohistochemistry (Paraffin) Western Blotting
Application Notes	Immunocytochemistry Analysis: 1:10-1:500 dilution of a previous lot was used. Neurons in culture should be permeablized with 0.1% triton X-100. All primary antibody dilutions should be performed with simple solutions containing only buffer and primary antibody without excess protein blocks or detergents. For dual labeling studies using mouse monoclonals, antibody incubations should be sequential with MAB377B last. First mouse monoclonal antibody should be first detected with anti-mouse secondary prior to incubating with MAB377B. Excess anti-mouse IgG may be blocked by incubating with 1% mouse serum prior to MAB377B incubation. Detection of biotinylated NeuN monoclonal is via streptavidin. In some cases in may be necessary to pretreat the tissue with avidin to block excess biotin prior to immunohistochemisty (Wood and Warnke, 1981). Immunohistochemistry: 1:200-1:2,000. The antibody works best on polyester wax embedded tissue but also works on paraffin embedded tissue at a lower working dilution. The antibody works well with formaldehyde-based fixatives. Citric acid and microwave pretreatment has been used successfully (Sarnat, 1998). Western Blotting Analysis: A previous lot of this antibody was used on western blot. Recognizes 2-3 bands in the 46-48 kDa range and possibly another band at approximately 66 kDa. Optimal working dilutions must be determined by end user.

Applications

Application

This Anti-NeuN Antibody, clone A60, biotin conjugated is validated for use in IC, IH, IH(P), WB for the detection of NeuN.

Key Applications

Immunocytochemistry
Immunohistochemistry
Immunohistochemistry (Paraffin)
Western Blotting

Application Notes

Immunocytochemistry Analysis:
1:10-1:500 dilution of a previous lot was used. Neurons in culture should be permeablized with 0.1% triton X-100. All primary antibody dilutions should be performed with simple solutions containing only buffer and primary antibody without excess protein blocks or detergents.

For dual labeling studies using mouse monoclonals, antibody incubations should be sequential with MAB377B last. First mouse monoclonal antibody should be first detected with anti-mouse secondary prior to incubating with MAB377B. Excess anti-mouse IgG may be blocked by incubating with 1% mouse serum prior to MAB377B incubation. Detection of biotinylated NeuN monoclonal is via streptavidin. In some cases in may be necessary to pretreat the tissue with avidin to block excess biotin prior to immunohistochemisty (Wood and Warnke, 1981).

Immunohistochemistry:
1:200-1:2,000. The antibody works best on polyester wax embedded tissue but also works on paraffin embedded tissue at a lower working dilution. The antibody works well with formaldehyde-based fixatives. Citric acid and microwave pretreatment has been used successfully (Sarnat, 1998).

Western Blotting Analysis:
A previous lot of this antibody was used on western blot. Recognizes 2-3 bands in the 46-48 kDa range and possibly another band at approximately 66 kDa.

Optimal working dilutions must be determined by end user.

Biological Information
Immunogen	Purified cell nuclei from mouse brain
Clone	A60
Concentration	Please refer to the Certificate of Analysis for the lot-specific concentration.
Host	Mouse
Specificity	Vertebrate neuron-specific nuclear protein called NeuN (Neuronal Nuclei). MAB377B reacts with most neuronal cell types throughout the nervous system of mice including cerebellum, cerebral cortex, hippocampus, thalamus, spinal cord and neurons in the peripheral nervous system including dorsal root ganglia, sympathetic chain ganglia and enteric ganglia. The immunohistochemical staining is primarily in the nucleus of the neurons with lighter staining in the cytoplasm. The few cell types not reactive with MAB377B include Purkinje, mitral and photoreceptor cells. Developmentally, immunoreactivity is first observed shortly after neurons have become postmitotic, no staining has been observed in proliferative zones. The antibody is an excellent marker for neurons in primary cultures and in retinoic acid-stimulated P19 cells. It is also useful for identifying neurons in transplants.
Isotype	IgG1
Species Reactivity	Mouse Rat Human Ferret Chicken Salamander
Species Reactivity Note	Rat and mouse. It is expected that the biotin conjugated antibody will also react with human, ferret, chick and salamander.
Antibody Type	Monoclonal Antibody
Gene Symbol	NeuN A59
Purification Method	Protein A Purfied
Molecular Weight	46-48 kDa

Biological Information

Immunogen

Purified cell nuclei from mouse brain

Clone

A60

Concentration

Please refer to the Certificate of Analysis for the lot-specific concentration.

Host

Mouse

Specificity

Vertebrate neuron-specific nuclear protein called NeuN (Neuronal Nuclei). MAB377B reacts with most neuronal cell types throughout the nervous system of mice including cerebellum, cerebral cortex, hippocampus, thalamus, spinal cord and neurons in the peripheral nervous system including dorsal root ganglia, sympathetic chain ganglia and enteric ganglia. The immunohistochemical staining is primarily in the nucleus of the neurons with lighter staining in the cytoplasm. The few cell types not reactive with MAB377B include Purkinje, mitral and photoreceptor cells. Developmentally, immunoreactivity is first observed shortly after neurons have become postmitotic, no staining has been observed in proliferative zones. The antibody is an excellent marker for neurons in primary cultures and in retinoic acid-stimulated P19 cells. It is also useful for identifying neurons in transplants.

Isotype

IgG1

Species Reactivity

Mouse
Rat
Human
Ferret
Chicken
Salamander

Species Reactivity Note

Rat and mouse. It is expected that the biotin conjugated antibody will also react with human, ferret, chick and salamander.

Antibody Type

Monoclonal Antibody

Gene Symbol

NeuN
A59

Purification Method

Protein A Purfied

Molecular Weight

46-48 kDa

Product Usage Statements
Quality Assurance	Routinely evaluated by immunohistochemistry on brain tissue. Immunohistochemistry(paraffin) Analysis: NeuN (cat. # MAB377B) staining pattern/morphology in rat cerebellum. Tissue pretreated with Citrate, pH 6.0. This lot of antibody was diluted to 1:100, using IHC-Select® Detection with HRP-DAB. Immunoreactivity is seen as nuclear staining in the neurons in the granular layer. Note that there is no signal detected in the nucleus of Purkinje cells. Optimal Staining With Citrate Buffer, pH 6.0, Epitope Retrieval: Rat Cerebellum
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Quality Assurance

Routinely evaluated by immunohistochemistry on brain tissue.

Immunohistochemistry(paraffin) Analysis:
NeuN (cat. # MAB377B) staining pattern/morphology in rat cerebellum. Tissue pretreated with Citrate, pH 6.0. This lot of antibody was diluted to 1:100, using IHC-Select® Detection with HRP-DAB. Immunoreactivity is seen as nuclear staining in the neurons in the granular layer. Note that there is no signal detected in the nucleus of Purkinje cells.
Optimal Staining With Citrate Buffer, pH 6.0, Epitope Retrieval: Rat Cerebellum

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Stable for 1 year at 2-8ºC from date of receipt.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at 2-8ºC from date of receipt.

Packaging Information
Material Size	500 µg

Packaging Information

Material Size

500 µg

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

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Ms X Neuronal Nuclei (NeuN), Biotin Conjugated

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