MANF Antibody [Unconjugated]

Western blot shows lysates of SH‑SY5Y human neuroblastoma cell line, U‑87 MG human glioblastoma/astrocytoma cell line, and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human MANF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3748) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for MANF at approximately 17 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Simple Western lane view shows lysates of SH‑SY5Y human neuroblastoma cell line and U‑87 MG human glioblastoma/astrocytoma cell line, loaded at 0.2 mg/mL. A specific band was detected for MANF at approximately 24 kDa (as indicated) using 10 µg/mL of Goat Anti-Human MANF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3748) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western blot shows lysates of HEK293T human embryonic kidney parental cell line and MANF knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human MANF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3748) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for MANF at approximately 17 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293Tcell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

• Reactivity: Hu
• Applications: WB, Simple Western, KO
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

MANF Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human MANF
  Leu22-Leu179
  Accession # P55145
• Specificity: Detects human MANF in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: MANF
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Knockout Validated
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for MANF Antibody [Unconjugated]

• Arginine-Rich Protein
• arginine-rich, mutated in early stage tumors
• ARMET
• ARP
• MANF
• mesencephalic astrocyte-derived neurotrophic factor
• Protein ARMET

Background

Mesencephalic astrocyte-derived neurotrophic factor (MANF), also known as arginine-rich, mutated in early stage tumors (ARMET) and arginine-rich protein (ARP), is a 20 kDa member of the ARMET family of proteins (1). The name ARMET comes from the fact that the protein was initially thought to be 50 aa longer at the N‑terminus and to contain an arginine-rich region (2-5). The presence of a specific mutation changing the previously numbered codon 50 from ATG to AGG, or deletion of that codon, has been reported in a variety of solid tumors (2-4). Human MANF is synthesized as a 179 amino acid (aa) precursor that contains a 21 aa signal sequence and a 158 aa mature chain. Mature human MANF is 99%, 98%, and 96% aa identical to mature rat, mouse and bovine MANF, respectively. MANF is localized to the endoplasmic reticulum (ER) and Golgi apparatus, and is also secreted (5).  In the CNS, MANF selectively protects nigral dopaminergic neurons, versus GABAergic or serotonergic neurons, which suggests that MANF may be indicated for the treatment of Parkinson’s disease (1). MANF is also one of the 12 commonly unfolded protein response (UPR)-upregulated genes (5). One study showed that MANF plays an important role in protecting cells against tunicamycin and thapsigargin-induced cell death (5). Loss of MANF renders cells more susceptible to those drugs, but also increases cell proliferation and decreases cell size (5). Another study showed that MANF is an endoplasmic reticulum stress response (ERSR) gene in the heart that can be induced and secreted in response to ER stresses, including ischemia, and that extracellular MANF may protect cardiac myocytes in an autocrine and paracrine manner (6). 

1. Petrova, P.S. et al. (2003) J. Mol. Neurosci. 20:173.
2. Shridhar, V. et al. (1996) Oncogene 12:1931.
3. Shridhar, R. et al. (1996) Cancer Res. 56:5576.
4. Shridhar, V. et al. (1997) Oncogene 14:2213.
5. Apostolou, A. et al. (2008) Exp. Cell Res. 314:2454.
6. Tadimalla, A. et al. (2008) Circ. Res. 103:1249.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for MANF Antibody (AF3748)(10)

Publications using AF3748

• Anttila, JE;Mattila, OS;Liew, HK;Mätlik, K;Mervaala, E;Lindholm, P;Lindahl, M;Lindsberg, PJ;Tseng, KY;Airavaara, M; MANF protein expression is upregulated in immune cells in the ischemic human brain and systemic recombinant MANF delivery in rat ischemic stroke model demonstrates anti-inflammatory effects Acta neuropathologica communications 2024-01-16 [PMID: 38229173] (ELISA Capture, Rat, Mouse)
• V Kovaleva, LY Yu, L Ivanova, O Shpironok, J Nam, A Eesmaa, EP Kumpula, S Sakson, U Toots, M Ustav, JT Huiskonen, MH Voutilaine, P Lindholm, M Karelson, M Saarma MANF regulates neuronal survival and UPR through its ER-located receptor IRE1alpha Cell Reports, 2023-02-03;42(2):112066. 2023-02-03 [PMID: 36739529] (Immunoprecipitation, Western Blot, Human)
• A Eesmaa, LY Yu, H Göös, T Danilova, K Nõges, E Pakarinen, M Varjosalo, M Lindahl, P Lindholm, M Saarma CDNF Interacts with ER Chaperones and Requires UPR Sensors to Promote Neuronal Survival International Journal of Molecular Sciences, 2022-08-22;23(16):. 2022-08-22 [PMID: 36012764] (ELISA Capture, Human, Mouse)
• J Konovalova, D Gerasymchu, SN Arroyo, S Kluske, F Mastroiann, AV Pereyra, A Domanskyi Human-Specific Regulation of Neurotrophic Factors MANF and CDNF by microRNAs International Journal of Molecular Sciences, 2021-09-07;22(18):. 2021-09-07 [PMID: 34575854] (ELISA Capture, Human)
• Tatiana Danilova, Emilia Galli, Emmi Pakarinen, Erik Palm, Päivi Lindholm, Mart Saarma et al. Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF) Is Highly Expressed in Mouse Tissues With Metabolic Function Front Endocrinol (Lausanne) 11/6/2019 [PMID: 31781038]
• E Galli, J Rossi, T Neumann, JO Andressoo, S Drinda, P Lindholm Mesencephalic Astrocyte-Derived Neurotrophic Factor Is Upregulated with Therapeutic Fasting in Humans and Diet Fat Withdrawal in Obese Mice Sci Rep, 2019-10-04;9(1):14318. 2019-10-04 [PMID: 31586115] (ELISA Capture, Human, Mouse)
• E Galli, A Planken, L Kadastik-E, M Saarma, P Taba, P Lindholm Increased Serum Levels of Mesencephalic Astrocyte-Derived Neurotrophic Factor in Subjects With Parkinson's Disease Front Neurosci, 2019-09-04;13(0):929. 2019-09-04 [PMID: 31555085] (ELISA Capture, Human)
• Emilia Galli, Taina Härkönen, Markus T. Sainio, Mart Ustav, Urve Toots, Arto Urtti et al. Increased circulating concentrations of mesencephalic astrocyte-derived neurotrophic factor in children with type 1 diabetes Scientific Reports 6/30/2016 [PMID: 27356471]
• Glembotski CC, Thuerauf DJ, Huang C Mesencephalic Astrocyte-derived Neurotrophic Factor Protects the Heart from Ischemic Damage and Is Selectively Secreted upon Sarco/endoplasmic Reticulum Calcium Depletion. J. Biol. Chem., 2012-05-25;287(31):25893-904. 2012-05-25 [PMID: 22637475] (Western Blot, Human)
• Tadimalla A, Belmont PJ, Thuerauf DJ, Glassy MS, Martindale JJ, Gude N, Sussman MA, Glembotski CC Mesencephalic astrocyte-derived neurotrophic factor is an ischemia-inducible secreted endoplasmic reticulum stress response protein in the heart. Circ. Res., 2008-10-16;103(11):1249-58. 2008-10-16 [PMID: 18927462] (ICC, Western Blot, Rat)

Bioinformatics

• Gene Symbol: MANF
• Uniprot:
  • P55145(Human)