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Mouse monoclonal antibody to proNGF (30-41), [Clone BS312]: IgG

$297.00USD


Catalogue No. M-1738-100
Description Nerve growth factor (NGF) is synthesized as a precursor (proNGF) which may be released and have physiological functions to cause cell death. It binds neurotrophin receptor p75 and sortilin and may also be important for the development of nervous system. proNGF is synthesized in target tissues and glia, transported retrogradely and may be released.
Related products Biotinylated proNGF antibody, cat# M-1778-50-B
Batch No. See product label
Unit size 100 ug
Antigen A synthetic peptide (C-HTIPQAHWTKLQ, aa: 30-41) of human proNGF protein has been used as the immunogen. The sequence is located on the pro-domain of the proNGF full-length protein and is 80% homologous to mouse and rat proNGF.
Antibody Type Mouse monoclonal IgG antibody
Isotype IgG2b, lambda-light chain
Other Names Pro-brain nerve growth factor; proNGF; NGF
Accession NGF_HUMAN
Produced in Mouse
Purity Protein G purified mouse IgG.
Applications Flow Cytometry (2 ug/ 10^6 cells). Immunocytochemistry (1-2 ug/mL), Western Blotting (1-2 ug/mL). Other applications not yet tested. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
Specificity Human
Cross-reactivity Species cross-reactivity not tested.
Form Lyophilised from a solution containing PBS buffer pH 7.4 with 3% trehalose, without preservatives.
Reconstitution Spin vial briefly before opening. Reconstitute in 100 uL of sterile water. Centrifuge to remove any insoluble material. Final buffer contains no preservatives.
Storage Store lyophilized antibody at 2-8C. After reconstitution divide into aliquots and store at -20C for a higher stability. Antibody contains no preservatives. Store at 2-8C with an appropriate antibacterial agent. Use sterile methods. Highest purity Glycerol (1:1) may be added for an additional stability when stored at refrigerated or freezing temperatures. Avoid repetitive freeze/thaw cycles.
Expiry Date 12 months after purchase if unopened
Images (click to zoom)
Mouse monoclonal antibody to proNGF (30-41), [Clone BS312]: IgG Fixing and Permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 200 ug/mL Normal Sheep IgG (20 minutes), Primary antibody: Mouse Monoclonal antibody to proNGF (cat # M-1738-100, 2 ug per ~10^6 cells) for 60 minutes at room temperature, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 fold dilution) with incubation for 20 minutes in dark at room temperature. Non-specific Control IgG, clone X63 (cat # M-1249-100) was used as negative control under same conditions (black dashed). Flow cytometry data and results were generated using Orflo MoxiflowTM instrument and protocols. The data demonstrates specific staining of proNGF expressed in human prostate cancer DU145 cell line using cat # M-1738-100.
Mouse monoclonal antibody to proNGF (30-41), [Clone BS312]: IgG Western blot analysis of proNGF expression in human brain homogenates. Monoclonal proNGF antibody M-1738-100 (2 μg/mL) detects monomeric, non-glycosylated proNGF protein (30 ng, E.coli derived) at 25 kDa (Sample 1). Brain homogenate 1 (Sample 2, 50 µg) and 2 (Sample 3, 30 µg) show several proNGF isoforms (~53 kDa, ~60 kDa, and ~73 kDa), which most likely relates to glycosylated forms of full-length proNGF (Reinshagen et al., 2000; Lobos et al., 2005; Pedraza et al., 2005; Pundavela et al., 2014). Specificity of these bands is shown by pre-incubation with the immunizing peptide (5x mass of primary antibody). Western Blotting Method: SDS-PAGE: denaturing and reducing, 4-20% Bis-Tris gel; Semi-Dry Transfer: Tris-Glycine (Towbin’s) buffer with 20% methanol; Membrane: Nitrocellulose (0.45 μm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C; Secondary antibody: anti-mouse-HRP (1/6000), 1 hour at RT; Detection: Chemiluminiscence.
Mouse monoclonal antibody to proNGF (30-41), [Clone BS312]: IgG Immunofluorescence analysis of proNGF expression in human DU145 prostate cancer cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) DU145 cells were incubated with proNGF antibody M-1738-100 (2 ug/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 ug/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). Magnification: 100x.
Mouse monoclonal antibody to proNGF (30-41), [Clone BS312]: IgG Western blot analysis of proNGF expression in DU145 and PC3 prostate cancer cell lysates. Monoclonal proNGF antibody M-1738-100 detects monomeric, non-glycosylated proNGF protein (20 ng, E.coli derived) at 25 kDa (Sample 1). PC3 (Sample 2, 100 µg) and DU145 (Sample 3, 100 µg) cell lysates show several proNGF isoforms (32 kDa, ~40 kDa, ~60 kDa, and higher molecular weight bands) as previously described, which most likely relates to glycosylated forms of full-length proNGF (Reinshagen et al., 2000; Lobos et al., 2005; Pedraza et al., 2005; Pundavela et al., 2014). Specificity of these bands is shown by pre-incubation with the immunizing peptide (5x mass of primary antibody), and a secondary antibody only control blot. Western Blotting Method: SDS-PAGE: denaturing and reducing, 4-20% Bis-Tris gel; Semi-Dry Transfer: Tris-Glycine (Towbin’s) buffer with 20% methanol; Membrane: Nitrocellulose (0.45 μm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: 2 ug/mL, overnight at 4°C; Secondary antibody: anti-mouse-HRP (1/6000), 1 hour at RT; Detection: Chemiluminiscence.
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