Anti-Brain-derived neurotrophic factor (BDNF) Antibody

Our Anti-Brain-derived neurotrophic factor (BDNF) rabbit polyclonal primary antibody detects human and other mammals (predicted) Brain-derived neurotrophic factor (BDNF), and is affinity-purified. It is validated for use in ELISA.

Western blot analysis of BDNF expression in human SH-SY5Y cell lysates (A) and human brain (B). Polyclonal rhBDNF antibody R-1707-100 (1 µg/mL) detects monomeric BDNF at 14 kDa, and monomeric proBDNF at 32 kDa in lysates prepared either in RIPA buffer or in acid-extraction buffer (A). Control antibody M-1744-50/100 (1 µg/mL) confirms detection of mature BDNF (Lane 1) and proBDNF (Lane 2) using BDNF and proBDNF proteins. A second band at 18 kDa is observed in the proBDNF standard, likely to represent a proBDNF degradation product. In human brain (B), R-1707-100 detects mature BDNF (14 kDa) and proBDNF (32 kDa) in both preparations (Tris-buffer and acid-treated). Acid-treated brain demonstrates lower background staining and gives a stronger BDNF signal.Western Blotting Method: SDS-PAGE: denaturing and reducing, 12% Bis-Tris gel; Transfer: Tris-Glycine buffer, semi-dry transfer; Membrane: nitrocellulose (0.22 µm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C; Secondary antibody: anti-mouse-HRP or anti-rabbit-HRP (1/6000), 2 hours at RT; Detection: Chemiluminiscence.
Analysis of BDNF expression in rat C6 cells by Flow Cytometry. Fixing and permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 200 µg/mL sheep IgG, Primary antibody: Rabbit polyclonal antibody to rhBDNF (cat # R-1707-100, 2 µg per ~10^6 cells) for 30 minutes at room temperature, Secondary antibody: Goat anti-rabbit-PE labeled secondary antibody (1:100 dilution), 20 minutes in dark at room temperature. Negative control: Normal rabbit IgG. Data and results were generated using Orflo MoxiflowTM instrument and protocols.
Western blot analysis of BDNF expression in human SH-SY5Y cell lysates (A) and human brain (B). Polyclonal rhBDNF antibody R-1707-100 (1 µg/mL) detects monomeric BDNF at 14 kDa, and monomeric proBDNF at 32 kDa in lysates prepared either in RIPA buffer or in acid-extraction buffer (A). Control antibody M-1744-50/100 (1 µg/mL) confirms detection of mature BDNF (Lane 1) and proBDNF (Lane 2) using BDNF and proBDNF proteins. A second band at 18 kDa is observed in the proBDNF standard, likely to represent a proBDNF degradation product. In human brain (B), R-1707-100 detects mature BDNF (14 kDa) and proBDNF (32 kDa) in both preparations (Tris-buffer and acid-treated). Acid-treated brain demonstrates lower background staining and gives a stronger BDNF signal.Western Blotting Method: SDS-PAGE: denaturing and reducing, 12% Bis-Tris gel; Transfer: Tris-Glycine buffer, semi-dry transfer; Membrane: nitrocellulose (0.22 µm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C; Secondary antibody: anti-mouse-HRP or anti-rabbit-HRP (1/6000), 2 hours at RT; Detection: Chemiluminiscence.
Western blot analysis of BDNF expression in human SH-SY5Y cell lysates (A) and human brain (B). Polyclonal rhBDNF antibody R-1707-100 (1 µg/mL) detects monomeric BDNF at 14 kDa, and monomeric proBDNF at 32 kDa in lysates prepared either in RIPA buffer or in acid-extraction buffer (A). Control antibody M-1744-50/100 (1 µg/mL) confirms detection of mature BDNF (Lane 1) and proBDNF (Lane 2) using BDNF and proBDNF proteins. A second band at 18 kDa is observed in the proBDNF standard, likely to represent a proBDNF degradation product. In human brain (B), R-1707-100 detects mature BDNF (14 kDa) and proBDNF (32 kDa) in both preparations (Tris-buffer and acid-treated). Acid-treated brain demonstrates lower background staining and gives a stronger BDNF signal.

Western Blotting Method: SDS-PAGE: denaturing and reducing, 12% Bis-Tris gel; Transfer: Tris-Glycine buffer, semi-dry transfer; Membrane: nitrocellulose (0.22 µm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C; Secondary antibody: anti-mouse-HRP or anti-rabbit-HRP (1/6000), 2 hours at RT; Detection: Chemiluminiscence.
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Human
ELISA, Flow, WB
Rabbit

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Data Sheet

SKU: R-1707-100

Volume/Size: 100 µg
Price:
$327.00
Technical Note 4 - Acid Extraction of Brain Tissue

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