Human MANF Antibody

R&D Systems, part of Bio-Techne | Catalog # AF3748

R&D Systems, part of Bio-Techne
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AF3748
AF3748-SP
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat

Applications

Validated:

Knockout Validated, Simple Western, Western Blot

Cited:

ELISA Capture, Immunocytochemistry, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all MANF Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human MANF
Leu22-Leu179
Accession # P55145

Specificity

Detects human MANF in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human MANF Antibody

Detection of Human MANF by Western Blot.

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.
Detection of Human MANF antibody by Simple WesternTM.

Detection of Human MANF by Simple WesternTM.

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 Human MANF Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human MANF Specificity by Using Knockout Cell Line.

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.

Applications for Human MANF Antibody

Application
Recommended Usage

Knockout Validated

MANF is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in MANF knockout HEK293T cell line.

Simple Western

10 µg/mL
Sample: SH‑SY5Y human neuroblastoma cell line and U‑87 MG human glioblastoma/astrocytoma cell line

Western Blot

1 µg/mL
Sample: SH‑SY5Y human neuroblastoma cell line, U‑87 MG human glioblastoma/astrocytoma cell line, and Daudi human Burkitt's lymphoma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 3 reviews rated 4 using AF3748 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Background: MANF

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). 

References

  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.

Long Name

Mesencephalic Astrocyte-derived Neurotrphic Factor

Alternate Names

ARMET, ARP

Entrez Gene IDs

7873 (Human); 74840 (Mouse); 315989 (Rat)

Gene Symbol

MANF

UniProt

P55145 (Human)

Additional MANF Products

Product Documents for Human MANF Antibody

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Product Specific Notices for Human MANF Antibody

For research use only

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