Invitrogen

NF1 Monoclonal Antibody (McNFn27b)

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NF1 Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on cancer biopsies of deparaffinized Human breast carcinoma tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and probed with a Neurofibromin monoclonal antibody (Product # MA1-085) at a dilution of 1:20 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

NF1 Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on normal biopsies of deparaffinized Human stomach tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and probed with a Neurofibromin monoclonal antibody (Product # MA1-085) at a dilution of 1:20 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

NF1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Neurofibromin in NIH-3T3 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a Neurofibromin monoclonal antibody (Product # MA1-085) at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). Neurofibromin staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

NF1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Neurofibromin was done on 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Neurofibromin (McNFn27b) Mouse Monoclonal Antibody (Product # MA1-085) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing punctate cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

NF1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Neurofibromin in U251 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a Neurofibromin monoclonal antibody (Product # MA1-085) at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). Neurofibromin staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

NF1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Neurofibromin in Hela cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a Neurofibromin monoclonal antibody (Product # MA1-085) at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). Neurofibromin staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1-3 µg/mL

-

Immunohistochemistry (IHC)

5-10 µg/mL

-

Immunocytochemistry (ICC/IF)

1-2 µg/mL

-

Product Specifications

Species Reactivity

Human, Mouse, Rat

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

McNFn27b

Immunogen

Synthetic peptide corresponding to the N-terminal residues 27-41 of human neurofibromin.

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein G

Storage buffer

PBS with 1mg/mL BSA

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_560293

Product Specific Information

MA1-085 detects neurofibromin in human, rat, and mouse cells.

MA1-085 has been successfully used in Western blot, and immunohistochemistry procedures. By Western blot, this antibody detects a ~240-260 kDa protein.

The MA1-085 immunogen is a synthetic peptide corresponding to the N-terminal residues 27-41 of human neurofibromin.

Target Information

The protein encoded by this gene is a type I integral membrane protein localized in the intermediate region between the endoplasmic reticulum and the Golgi, presumably recycling between the two compartments. The protein is a mannose-specific lectin and is a member of a novel family of plant lectin homologs in the secretory pathway of animal cells. Mutations in the gene are associated with a coagulation defect. Using positional cloning, the gene was identified as the disease gene leading to combined factor V-factor VIII deficiency, a rare, autosomal recessive disorder in which both coagulation factors V and VIII are diminished.

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

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Bioinformatics

Protein Aliases: DKFZp686J1293; FLJ21220; neurofibromatosis 1; Neurofibromatosis type 1; Neurofibromatosis-related protein NF-1; Neurofibromin; Neurofibromin truncated

Gene Aliases: AW494271; E030030H24Rik; Nf-1; NF1; NFNS; VRNF; WSS

UniProt ID: (Human) P21359, (Mouse) Q04690, (Rat) P97526

Entrez Gene ID: (Human) 4763, (Mouse) 18015, (Rat) 24592

Molecular Function: G-protein modulator enzyme modulator

MAPK cascade osteoblast differentiation metanephros development response to hypoxia liver development negative regulation of endothelial cell proliferation positive regulation of endothelial cell proliferation regulation of cell-matrix adhesion negative regulation of cell-matrix adhesion negative regulation of protein kinase activity cell communication Ras protein signal transduction negative regulation of neuroblast proliferation brain development peripheral nervous system development heart development skeletal muscle tissue development visual learning extrinsic apoptotic signaling pathway via death domain receptors regulation of gene expression Schwann cell development phosphatidylinositol 3-kinase signaling negative regulation of angiogenesis spinal cord development amygdala development forebrain astrocyte development neural tube development cerebral cortex development myelination in peripheral nervous system actin cytoskeleton organization extracellular matrix organization collagen fibril organization adrenal gland development negative regulation of cell migration regulation of synaptic transmission, GABAergic cellular response to heat negative regulation of Rac protein signal transduction wound healing negative regulation of transcription factor import into nucleus positive regulation of apoptotic process regulation of GTPase activity negative regulation of MAP kinase activity negative regulation of MAPK cascade pigmentation positive regulation of neuron apoptotic process regulation of blood vessel endothelial cell migration positive regulation of GTPase activity regulation of bone resorption negative regulation of osteoclast differentiation regulation of glial cell differentiation positive regulation of adenylate cyclase activity regulation of angiogenesis negative regulation of Ras protein signal transduction negative regulation of neurotransmitter secretion negative regulation of fibroblast proliferation regulation of long-term neuronal synaptic plasticity sympathetic nervous system development camera-type eye morphogenesis negative regulation of astrocyte differentiation negative regulation of oligodendrocyte differentiation smooth muscle tissue development artery morphogenesis forebrain morphogenesis cognition gamma-aminobutyric acid secretion, neurotransmission glutamate secretion, neurotransmission observational learning regulation of long-term synaptic potentiation positive regulation of extrinsic apoptotic signaling pathway via death domain receptors positive regulation of extrinsic apoptotic signaling pathway in absence of ligand signal transduction negative regulation of cell proliferation regulation of cell proliferation regulation of MAPK cascade regulation of neuron differentiation

Process(es)

GTPase activator activity protein binding phosphatidylethanolamine binding phosphatidylcholine binding microtubule binding lipid binding syndecan binding