Human NRG1 Isoform SMDF Antibody

Catalog # Availability Size / Price Qty
AF378
AF378-SP

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Cell Proliferation Induced by NRG1 Isoform SMDF and Neutralization by Human NRG1 Isoform SMDF Anti-body.
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Human NRG1 Isoform SMDF Antibody Summary

Species Reactivity
Human
Specificity
Detects human NRG1 Isoform SMDF in direct ELISAs and Western blots. Neutralizes 60‑80% of the biological activity of human NRG1 Isoform SMDF.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human NRG1 Isoform SMDF
Met1-Glu296
Accession # NP_039253
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.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human NRG1 Isoform SMDF (Catalog # 378-SM)
Neutralization
Measured by its ability to neutralize NRG1 Isoform SMDF-induced proliferation in the MCF‑7 human breast cancer cell line. Karey, K. P. et al. (1988) Cancer Research 48:4083. The Neutralization Dose (ND50) is typically 2‑8 µg/mL in the presence of 12 ng/mL Recombinant Human NRG1 Isoform SMDF.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Neutralization Cell Proliferation Induced by NRG1 Isoform SMDF and Neutralization by Human NRG1 Isoform SMDF Anti-body. View Larger

Cell Proliferation Induced by NRG1 Isoform SMDF and Neutralization by Human NRG1 Isoform SMDF Anti-body. Recombinant Human NRG1 Isoform SMDF (Catalog # 378-SM) stimulates proliferation in the MCF-7 human breast cancer cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human NRG1 Isoform SMDF (12 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human NRG1 Isoform SMDF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF378). The ND50 is typically 2-8 µg/mL.

Western Blot Detection of Neuregulin-1 Isoform SMDF by Western Blot View Larger

Detection of Neuregulin-1 Isoform SMDF by Western Blot SMDF transforms via ErbB receptor signalling.(A) Western blot analysis of indicated proteins in NSLT cells expressing SMDF, SMDF* (point mutant incapable of binding ErbB receptors) or control vector (LXSN), and SMDF(T) cells derived from SMDF tumours in the absence or presence of the ErbB-2 inhibitor, 4557W (40 nM). (B) Cell counts of triplicate wells in the presence or absence of the indicated concentrations of 4557W −/+ S.D.. Figure is representative of 3 independent experiments. (C) Number of colonies formed in soft agar assays of NSLT cells expressing SMDF, Ras or control vector LXSN in the absence or presence of the indicated inhibitors. U0126 (UO), LY294002 (LY) and the ErbB inhibitor (ErbBI). 12 fields were counted per well with the results shown as average/well −/+ S.D.. Results are representative of two separate experiments (D) Soft agar assays of NSLT cells infected with SMDF, SMDF* or control vector (E) 105 NSLT cells expressing SMDF, Ras or control vector (LXSN) were seeded in triplicate into 6 well dishes and counted at 72 h. The inhibitors U0126 (20 µM), LY294002 (20 µM) or control vehicle were added where indicated at 36 hours. Western blot analysis of total lysates from NSLT cells expressing SMDF or control vector (LXSN). (F) Western blot analysis of total lysates from NSLT cells expressing SMDF or control vector (LXSN) collected 3 (preconfluent), 6 and 9 days after seeding. Plates were medium changed daily. (G) Western blot analysis of total lysates from NSLT cells expressing SMDF, Ras or control vector (LXSN) in attached (att) or anchorage-independent conditions (sus). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20668675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Neuregulin-1 Isoform SMDF by Western Blot View Larger

Detection of Neuregulin-1 Isoform SMDF by Western Blot SMDF transforms via ErbB receptor signalling.(A) Western blot analysis of indicated proteins in NSLT cells expressing SMDF, SMDF* (point mutant incapable of binding ErbB receptors) or control vector (LXSN), and SMDF(T) cells derived from SMDF tumours in the absence or presence of the ErbB-2 inhibitor, 4557W (40 nM). (B) Cell counts of triplicate wells in the presence or absence of the indicated concentrations of 4557W −/+ S.D.. Figure is representative of 3 independent experiments. (C) Number of colonies formed in soft agar assays of NSLT cells expressing SMDF, Ras or control vector LXSN in the absence or presence of the indicated inhibitors. U0126 (UO), LY294002 (LY) and the ErbB inhibitor (ErbBI). 12 fields were counted per well with the results shown as average/well −/+ S.D.. Results are representative of two separate experiments (D) Soft agar assays of NSLT cells infected with SMDF, SMDF* or control vector (E) 105 NSLT cells expressing SMDF, Ras or control vector (LXSN) were seeded in triplicate into 6 well dishes and counted at 72 h. The inhibitors U0126 (20 µM), LY294002 (20 µM) or control vehicle were added where indicated at 36 hours. Western blot analysis of total lysates from NSLT cells expressing SMDF or control vector (LXSN). (F) Western blot analysis of total lysates from NSLT cells expressing SMDF or control vector (LXSN) collected 3 (preconfluent), 6 and 9 days after seeding. Plates were medium changed daily. (G) Western blot analysis of total lysates from NSLT cells expressing SMDF, Ras or control vector (LXSN) in attached (att) or anchorage-independent conditions (sus). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20668675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Neuregulin-1 Isoform SMDF by Western Blot View Larger

Detection of Neuregulin-1 Isoform SMDF by Western Blot SMDF transforms via ErbB receptor signalling.(A) Western blot analysis of indicated proteins in NSLT cells expressing SMDF, SMDF* (point mutant incapable of binding ErbB receptors) or control vector (LXSN), and SMDF(T) cells derived from SMDF tumours in the absence or presence of the ErbB-2 inhibitor, 4557W (40 nM). (B) Cell counts of triplicate wells in the presence or absence of the indicated concentrations of 4557W −/+ S.D.. Figure is representative of 3 independent experiments. (C) Number of colonies formed in soft agar assays of NSLT cells expressing SMDF, Ras or control vector LXSN in the absence or presence of the indicated inhibitors. U0126 (UO), LY294002 (LY) and the ErbB inhibitor (ErbBI). 12 fields were counted per well with the results shown as average/well −/+ S.D.. Results are representative of two separate experiments (D) Soft agar assays of NSLT cells infected with SMDF, SMDF* or control vector (E) 105 NSLT cells expressing SMDF, Ras or control vector (LXSN) were seeded in triplicate into 6 well dishes and counted at 72 h. The inhibitors U0126 (20 µM), LY294002 (20 µM) or control vehicle were added where indicated at 36 hours. Western blot analysis of total lysates from NSLT cells expressing SMDF or control vector (LXSN). (F) Western blot analysis of total lysates from NSLT cells expressing SMDF or control vector (LXSN) collected 3 (preconfluent), 6 and 9 days after seeding. Plates were medium changed daily. (G) Western blot analysis of total lysates from NSLT cells expressing SMDF, Ras or control vector (LXSN) in attached (att) or anchorage-independent conditions (sus). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20668675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Neuregulin-1 Isoform SMDF by Western Blot View Larger

Detection of Neuregulin-1 Isoform SMDF by Western Blot SMDF promotes oncogenic transformation.(A) Western blot analysis of total lysates from NSLT cells selected to express human SMDF or control vector (LXSN). SMDF(T) are derived from tumours, induced by injecting NSLTSMDF cells into nude mice. (B) Cell counts −/+ S.D. following seeding of 105 NSLT cells per well expressing SMDF, Ras or control vector (LXSN) in triplicate in 6 well plates. Medium was changed daily. Experiment shown is representative of 3 separate experiments. Representative phase-contrast images of the cells at high density. (C) NSLT cells expressing SMDF, Ras or control vector (LXSN) were injected into the flanks of nude mice. Ras cells were only injected into a single flank. Picture shows examples of tumours formed (SMDF (left), Ras (right)) and the graph shows the average growth of the tumours −/+ S.D.. (D) Representative images of cells dissociated from NSLTSMDF derived tumours stained for LT (upper panel) or the Schwann cell marker, S100 (lower panel). Primary Schwann cells (NS) were infected with retroviral vectors expressing SMDF, Ras or the control vector (LXSN) and selected in G418. One week following selection, the cultures were fixed and stained for the senescence marker beta -Galactosidase at pH 6 (E) or seeded in triplicate and counted for four passages (F). Western blot shows p-ERK levels in lysates prepared from NS cells expressing Ras, SMDF or control vector (LXSN) six days after infection. (G) 2×105 NS cells infected with SMDF or control vector (LXSN) were plated in triplicate in 6 well dishes and counted at the indicated days. Medium was changed daily. Error bars show −/+ S.D.. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20668675), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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: Neuregulin-1 Isoform SMDF

The heregulin (also known as neuregulin) family of cytokines is comprised of multiple secreted or membrane-bound isoforms that are produced from the single heregulin gene by alternative splicing and/or usage of different promoters. All heregulin family members share an epidermal growth factor (EGF)-like domain ( alpha - or beta ‑variant) that interacts with the erbB family of tyrosine kinase receptors. NRG1 Isoform SMDF is a heregulin isoform containing a C-terminal EGF-like domain ( beta ‑variant) and a unique N-terminal sequence that lacks an Ig-like domain which is present in all other known heregulins. NRG1 Isoform SMDF also lacks a transmembrane domain and the cytoplasmic tail. NRG1 Isoform SMDF expression has been found to be restricted to the nervous system. It is likely that NRG1 Isoform SMDF may play an important role in neural-specific functions.

References
  1. Yarden, Y. and D. Wen (1994) in Guidebook to Cytokines and Their Receptors,
  2. N.A. Nicola, Ed., Oxford University Press, p. 146.
  3. Meyer, D. et al. (1997) Development 124:3575.
Entrez Gene IDs
3084 (Human); 211323 (Mouse); 112400 (Rat)
Alternate Names
ARIA; GGF; GGF2; HGL; HRG; HRG1; HRGA; MST131; MSTP131; NDF; Neuregulin1 Isoform SMDF; Neuregulin-1 Isoform SMDF; NRG1-IT2; SMDF

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