Invitrogen

NF-H Monoclonal Antibody (3G3)

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NF-H Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence of neurofilament, heavy chain in rat cerebral cortex cultures in green.

NF-H Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Neurofilament Heavy Chain showing staining in the cytoplasm and nucleus of paraffin-embedded rat brain tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Neurofilament Heavy Chain monoclonal antibody (Product # MA1-2012) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

NF-H Antibody in Western Blot (WB) — Western blot analysis of Neurofilament, Heavy chain was performed by loading 25 µg of SH-SY5Y (lane 1) and rat brain (lane 2) lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a Neurofilament, Heavy chain monoclonal antibody (Product # MA1-2012) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~200 kDa.

Product Details

MA1-2012

Applications	Tested Dilution	Publications
Western Blot (WB)	1:100-1:1,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:10-1:100	-
Immunocytochemistry (ICC/IF)	5 µg/mL	-
Immunoprecipitation (IP)	-	View 10 publications 10 publications

Product Specifications
Species Reactivity	Human, Rat
Published species	Fruit fly, Human, Mouse, Rabbit
Host/Isotype	Mouse
Class	Monoclonal
Type	Antibody
Clone	3G3
Immunogen	Synthetic peptide corresponding to 226 amino acids of the C-terminus of neurofilament, heavy chain. View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	purified
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_347005

Product Specific Information

MA1-2012 detects the neurofilament, heavy chain in human and rat samples.

MA1-2012 has been successfully used in Western blot, immunohistochemistry and immunofluorecence procedures. By Western blot this antibody detects a ~200 kDa protein representing the neurofilament, heavy chain in rat brain microsome. In immunohistochemistry procedures MA1-2012 recognizes the neurofilament, heavy chain in rat hippocampal neurons.

The MA1-2012 immunogen is a synthetic peptide corresponding to 226 amino acids of the C-terminus of neurofilament, heavy chain.

Target Information

Involved in the maintenance of neuronal caliber, neurofilaments are the intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H. Like most other intermediate filament proteins (IFPs), the expression of the different neuronal IFPs is both tissue-specific and developmentally regulated. NF-H has an important role in mature axons and is involved in amyotrophic lateral sclerosis (ALS).

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Protein phosphatase 5 is a major component of glucocorticoid receptor.hsp90 complexes with properties of an FK506-binding immunophilin.

The Journal of biological chemistry

Silverstein AM,Galigniana MD,Chen MS,Owens-Grillo JK,Chinkers M,Pratt WB

MA1-2012 was used in immunoprecipitation to identify PP5 as an important component of glucocorticoid receptor-hsp90 complexes and investigate its functional properties

Fri Jun 27 00:00:00 EDT 1997

Folding of the glucocorticoid receptor by the reconstituted Hsp90-based chaperone machinery. The initial hsp90.p60.hsp70-dependent step is sufficient for creating the steroid binding conformation.

The Journal of biological chemistry

Dittmar KD,Pratt WB

MA1-2012 was used in immunoprecipitation to study the folding of hormone binding domain of glucocorticoid receptor in vitro using reconstituted receptor-hsp90 heterocomplex assembly system with purified components

Fri May 16 00:00:00 EDT 1997

Functional interference between hypoxia and dioxin signal transduction pathways: competition for recruitment of the Arnt transcription factor.

Molecular and cellular biology

Gradin K,McGuire J,Wenger RH,Kvietikova I,fhitelaw ML,Toftgård R,Tora L,Gassmann M,Poellinger L

MA1-2012 was used in immunoprecipitation to show HIF-1a is associated with the molecular chaperone hsp90 in vitro

Tue Oct 01 00:00:00 EDT 1996

A model of protein targeting mediated by immunophilins and other proteins that bind to hsp90 via tetratricopeptide repeat domains.

The Journal of biological chemistry

Owens-Grillo JK,Czar MJ,Hutchison KA,Hoffmann K,Perdew GH,Pratt WB

MA1-2012 was used in immunoprecipitation to study the mechanisms of immunophilin-mediated protein targeting

Fri Jun 07 00:00:00 EDT 1996

The basic helix-loop-helix/PAS factor Sim is associated with hsp90. Implications for regulation by interaction with partner factors.

The Journal of biological chemistry

McGuire J,Coumailleau P,Whitelaw ML,Gustafsson JA,Poellinger L

MA1-2012 was used in immunoprecipitation to study the interaction between Sim and hsp90

Fri Dec 29 00:00:00 EST 1995

Identification of functional domains of the aryl hydrocarbon receptor.

The Journal of biological chemistry

Fukunaga BN,Probst MR,Reisz-Porszasz S,Hankinson O

MA1-2012 was used in immunoprecipitation to investigate the functional domains of mouse aryl hydrocarbon receptor

Fri Dec 08 00:00:00 EST 1995

Definition of a minimal domain of the dioxin receptor that is associated with Hsp90 and maintains wild type ligand binding affinity and specificity.

The Journal of biological chemistry

Coumailleau P,Poellinger L,Gustafsson JA,Whitelaw ML

MA1-2012 was used in immunoprecipitation to detect the interaction between the dioxin receptor and Hsp90

Fri Oct 20 00:00:00 EDT 1995

A tyrosine kinase-dependent pathway regulates ligand-dependent activation of the dioxin receptor in human keratinocytes.

The Journal of biological chemistry

Gradin K,Whitelaw ML,Toftgård R,Poellinger L,Berghard A

MA1-2012 was used in immunoprecipitation to investigate the role of a tyrosine kinase-dependent pathway during ligand-dependent activation of the dioxin receptor in human keratinocytes

Fri Sep 23 00:00:00 EDT 1994

Characterization of the protein-protein interactions determining the heat shock protein (hsp90.hsp70.hsp56) heterocomplex.

The Journal of biological chemistry

Czar MJ,Owens-Grillo JK,Dittmar KD,Hutchison KA,Zacharek AM,Leach KL,Deibel MR,Pratt WB

MA1-2012 was used in immunoprecipitation to investigate the interaction of different protein components of the heat shock protein heterocomplex

Fri Apr 15 00:00:00 EDT 1994

A cellular factor stimulates ligand-dependent release of hsp90 from the basic helix-loop-helix dioxin receptor.

Molecular and cellular biology

McGuire J,Whitelaw ML,Pongratz I,Gustafsson JA,Poellinger L

MA1-2012 was used in immunoprecipitation to investigate the role of Arnt for release of hsp90 from the dioxin receptor in the presence of dioxin

Fri Apr 01 00:00:00 EST 1994

Bioinformatics

Protein Aliases: 200 kDa neurofilament protein; NEF3; neurofilament; Neurofilament heavy polypeptide; Neurofilament triplet H protein; neurofilament, heavy polypeptide 200kDa; NF 200 kD; NF-H; NHC

Gene Aliases: CMT2CC; KIAA0845; NEFH; NFH

UniProt ID: (Human) P12036

Entrez Gene ID: (Human) 4744, (Rat) 24587

Function(s)

microtubule cytoskeleton organization ovarian follicle atresia axonogenesis spinal cord development hippocampus development cerebral cortex development cell projection assembly axon regeneration neurofilament bundle assembly cellular response to oxidative stress response to cocaine peripheral nervous system neuron axonogenesis axon development cellular response to estradiol stimulus regulation of organelle transport along microtubule response to sodium arsenite response to acrylamide brain development embryo development neuron projection development intermediate filament cytoskeleton organization intermediate filament bundle assembly neurofilament cytoskeleton organization

Process(es)

structural molecule activity structural constituent of cytoskeleton microtubule binding toxic substance binding kinesin binding protein kinase binding protein binding, bridging dynein binding protein heterodimerization activity

NF-H Monoclonal Antibody (3G3)

Product Details

MA1-2012

Western Blot (WB)

Immunohistochemistry (Paraffin) (IHC (P))

Immunocytochemistry (ICC/IF)

Immunoprecipitation (IP)

Species Reactivity

Published species

Host/Isotype

Class

Type

Clone

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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