MANF Antibody

• Catalog Number: orb1239620
• Category: Antibodies
• Description: MANF Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC-P, WB
• Predicted Reactivity: Bovine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Anti-MANF antibody (orb1239620) was raised against a peptide corresponding to 12 amino acids near the amino terminus of human MANF. The immunogen is located within the first 50 amino acids of MANF.
• Concentration: 1 mg/mL
• Dilution range: WB: 0.125-2 μg/mL; IHC: 2.5 μg/mL; IF: 20 μg/mL.Antibody validated: Western Blot in human and rat samples; Immunohistochemistry in human samples; Immunofluorescence in human samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 20kDObserved: 26 kD
• Target: MANF
• UniProt ID: P55145
• NCBI: P55145
• Storage: MANF antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: MANF Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: MANF Antibody: ARP, ARMET, ARP, Mesencephalic astr
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western Blot Validation in Human Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: MANF orb1239620, (2 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation with Recombinant Protein. Loading: 30 ng of human MANF recombinant protein per lane. Antibodies: MANF orb1239620 (Lane 1: 0.125 µg/mL, Lane 2: 0.25 µg/mL and Lane 3: 0.5 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Observed at around 55kD.

Western Blot Validation in Rat Brain Tissue Lysate. Loading: 15 µg of lysates per lane. Antibodies: MANF orb1239620 (A: 1 µg /mL and B: 2 µg /mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunohistochemistry Validation of MANF in Human Brain Tissue. Immunohistochemical analysis of paraffin-embedded human brain tissue using anti-MANF antibody (orb1239620) at 2.5 µg /ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunofluorescence Validation of MANF in Human Brain Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed human brain tissue labeling MANF with orb1239620 at 20 µg /mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Induced Expression Validation of MANF in Mouse C17.2 Neural Stem Cells (Almutawaa et al., 2014). Concentration- and time-dependent effects of Valproic acid (VPA) on MANF at 48 h examined by Western blot analysis with anti-MANF antibodies. MANF was markedly increased 48h after VPA treatment. Lanes 1–4: control, 0.5 mM VPA, 1 mM VPA, 3 mM VPA.