MANF Antibody

• Catalog Number: orb1239622
• Category: Tools
• Description: MANF Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC-P, WB
• Isotype: IgG
• Immunogen: MANF antibody was raised against a 12 amino acid synthetic peptide from near the carboxy terminus of human MANF.The immunogen is located within the last 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/mLAntibody validated: Western Blot in human, mouse 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: 18 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
• Application notes: WB: 0.125-2 μg/mL; IHC: 2.5 μg/mL; IF: 20 μg/mLAntibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human samples; Immunofluorescence in human samples. All other applications and species not yet tested.
• Expiration Date: 12 months from date of receipt.

KO Validation in HEK293T Cells. Loading: 10 µg of HEK293T WT cell lysates or MANF KO cell lysates. Antibodies: MANF orb1239622 (1 µg/mL) and beta-actin orb1240312 (1 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Human, Mouse (NIH/3T3) and Rat (YB2/0) Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: MANF orb1239622, (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 orb1239622 (Lane 1: 0.125 µg /mL, Lane 2: 0.25µg /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 Human Tissues. Loading: 15 µg of lysates per lane. Antibodies: MANF orb1239622, (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 in Mouse Tissues. Loading: 15 µg of lysates per lane. Antibodies: MANF orb1239622, (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 in Rat Liver. Loading: 15 µg of lysates per lane. Antibodies: MANF orb1239622, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 µg/mL, Lane 2: 2 µg/mL, Lane 3: 4 µg/mL.

Immunofluorescence Validation of MANF in Human Cervix. Immunofluorescent analysis of 4% paraformaldehyde-fixed human cervix cells labeling MANF with orb1239622 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).

Immunofluorescence Validation of MANF in Human Colon. Immunofluorescent analysis of 4% paraformaldehyde-fixed human colon cells labeling MANF with orb1239622 at 5 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).

Immunofluorescence Validation of MANF in Human Liver. Immunofluorescent analysis of 4% paraformaldehyde-fixed human liver cells labeling MANF with orb1239622 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).

Immunofluorescence Validation of MANF in Mouse Pancreas. Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse pancreas cells labeling MANF with orb1239622 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).

Immunofluorescence Validation of MANF in Rat Liver. Immunofluorescent analysis of 4% paraformaldehyde-fixed rat liver cells labeling MANF with orb1239622 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI antibody (blue).

Immunohistochemistry Validation of MANF in Human Brain Tissue. Immunohistochemical analysis of paraffin-embedded human brain tissue using anti-MANF antibody (orb1239622) 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.

Localization and Immunocytochemistry Validation of MANF in adult zebrafish brain. (Chen et al., 2012). Co-immunostaining with anti-MANF antibodies (orb1239622) shows that majority of MANF-positive cells were HuC/D positive cells (fig. C-E) whereas a few MANF-positive cells were either glial (zrf-1) positive cells (fig. F-H) or TH1 positive cells (fig I-K). It indicates that MANF was mostly expressed in mature neuronal cells. MANF staining is shown in green.

KD Validation of MANF in Zebrafish Embryo (Chen et al., 2012). Western blot analysis with anti-MANF antibodies (orb1239622) was performed for MANF in 3dpf zebrafish embryo. After injected with a combination of two antisense splice-blocking morpholino oligonucleotides (MOsp), MANF expression was reduced by 85% as compared to the wild type.

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.