MANF Antibody Cat. No.: 4347

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psi-iconSpecifications
HOST SPECIES:Rabbit
SPECIES REACTIVITY: Human, Mouse, Rat
HOMOLOGY: Predicted species reactivity based on immunogen sequence: Bovine: (92%)
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.
TESTED APPLICATIONS: ELISA, IF, IHC-P, WB
APPLICATIONS: MANF antibody can be used for detection of MANF by Western blot at 1 - 2 μg/mL. Antibody can also be used for immunohistochemistry starting at 2.5 μg/mL. For immunofluorescence start at 20 μg/mL.

Antibody validated: Western Blot in mouse samples; Immunohistochemistry in human samples and Immunofluorescence in human samples. All other applications and species not yet tested.
SPECIFICITY: This antibody does not cross-react with CDNF.
POSITIVE CONTROL:1) Cat. No. 1403 - Mouse Brain Tissue Lysate
 2) Cat. No. 10-301 - Human Brain Tissue Slide
 3) Cat. No. 1224 – Daudi Cell Lysate
 4) Cat. No. 1282 – 3T3/NIH Lysate
PREDICTED MOLECULAR WEIGHT: 20 kDa
psi-iconAdvanced Validation
VALIDATION:

KO Validation (Figure 1): Anti-MANF antibodies (4347) specificity was further verified by MANF specific knockout. MANF signal was not detected in MANF knockout HEK293T cells as compared to that in control wild type cells.

Recombinant Protein Test (Figure 3): Anti-MANF antibodies (4347) detected human MANF recombinant protein at different concentrations.

Localization Validation (Figure 7): MANF detected by anti-MANF antibodies is colocalized with HuC/D (a general neural marker).  MANF is localized in mature neuronal cells.

KD Validation (Figure 8): Anti-MANF antibody (4347) specificity was further verified by MANF specific knockdown. MANF signal, when injected with a combination of two antisense splice-blocking morpholino oligonucleotides (MOsp), was disrupted in comparison with either wild type or when injected with control MO .

Induced Expression Validation (Figure 9): MANF expression detected by anit-MANF antibodies was up-regulated by VPA treatment.

ISOFORMS: Human MANF has one isoform (182aa, 21kD). Mouse MANF has one isoform (179aa, 20kD) and Rat MANF also has one isoform (179aa, 20kD). 4347 can detect human, mouse and rat.

psi-iconProperties
PURIFICATION:MANF Antibody is affinity chromatography purified via peptide column.
CLONALITY:Polyclonal
ISOTYPE:IgG
CONJUGATE:Unconjugated
PHYSICAL STATE:Liquid
BUFFER:MANF Antibody is supplied in PBS containing 0.02% sodium azide.
CONCENTRATION:1 mg/mL
STORAGE CONDITIONS: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.

psi-iconAdditional Info
OFFICIAL SYMBOL:MANF
ALTERNATE NAMES:MANF Antibody: ARP, ARMET, ARP, Mesencephalic astrocyte-derived neurotrophic factor, Arginine-rich protein
ACCESSION NO.:P55145
PROTEIN GI NO.:23503040
GENE ID:7873
USER NOTE:Optimal dilutions for each application to be determined by the researcher.
psi-iconBackground and References
BACKGROUND:MANF Antibody: MANF, also known as ARMET, was initially identified as a protein containing an arginine-rich region that was highly mutated in a variety of tumors. More recently it was identified as a mesencephalic astrocyte-derived neurotrophic factor with selectivity for dopaminergic neurons, similar to glial cell line-derived neurotrophic factor (GDNF) and CDNF. In rat brain slices, MANF enhanced nigral gamma-aminobutyric acid release. Like GDNF and CDNF, MANF has selective neuroprotective activity for dopaminergic neurons suggesting that it may be indicated for the treatment of Parkinson's disease. Expression of MANF has also been shown to be induced during ER stress, suggesting that it may play a role in protein quality control during ER stress.
REFERENCES:1) Shridhar et al. Oncogene1996; 12:1931-9.
2) Shridhar et al. Cancer Res.1996; 56:5576-8.
3) Petrova et al. J. Mol. Neurosci.2003; 20:173-88.
4) Lindholm et al. Nature 2007; 448:73-7.
psi-iconCITATIONS
CITATIONS: 1)Chen et al. MANF regulates dopaminergic neuron development in larval zebrafish. Dev Biol. 2012;370(2):237-49. PMID: 22898306
2)Almutawaa et al. Induction of neurotrophic and differentiation factors in neural stem cells by valproic acid. Basic Clin Pharmacol Toxicol. 2014;115(2):216-21. PMID: 24460582
3)Niles et al. Valproic acid up-regulates melatonin MT1 and MT2 receptors and neurotrophic factors CDNF and MANF in the rat brain. Int J Neuropsychopharmacol. 2012;15(9):1343-50. PMID: 22243807
4)Walaa S. Almutawaa. Induction Of Neurotrophic and Differentiation Genes in Neural Stem Cells by Valproic Acid. McMaster University, M.Sc. Thesis. 2012.PMID:
5)Merja H. Voutilainen. CDNF and MANF in an experimental model of Parkinson's disease in rats. University of Helsinki, Ph.D. Thesis. 2010.PMID:

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