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Mouse monoclonal antibody to rh BDNF [4C8]: IgG

$227.00USD


Catalogue No. M-1744-50
Description BDNF belongs to the neurotrophin family and promotes the survival of neuronal populations that are all located either in the central nervous system or directly connected to it. It is a major regulator of synaptic transmission and plasticity at adult synapses in many regions of the CNS. The versatility of BDNF is emphasized by its contribution to a range of adaptive neuronal responses including long-term potentiation (LTP), long-term depression (LTD), certain forms of short-term synaptic plasticity, as well as homeostatic regulation of intrinsic neuronal excitability. The alterations in BDNF expression induced by various kinds of brain insult including stress, ischemia, seizure activity and hypoglycemia, may contribute to some pathologies such as depression, epilepsy, Alzheimer's, and Parkinson's disease. Microglia release BDNF that may contribute to neuroinflammation and neuropathic pain. SUBUNIT: Monomers and homodimers. Binds to NTRK2/TRKB. SUBCELLULAR LOCATION: Secreted protein. POst translation modification: Converted into mature BDNF by plasmin (PLG). SIMILARITY: Belongs to the NGF-beta family.
Related products Available as 100 g (2 x 50 g).
Batch No. See product label.
Unit size 50 g.
Antigen Recombinant human mature BDNF expressed in E.coli
Antibody Type Mouse monoclonal.
Clone 4C8
Other Names Brain-derived neurotrophic factor; Abrineurin; proBDNF
Accession BDNF_HUMAN
Produced in Mouse
Molecular Weight Mature BDNF dimer has a molecular weight of 28 kDa. In Western Blotting, mature BDNF monomer migrates at 14 kDa.
Purity Protein G purified mouse IgG.
Applications ELISA: 50-100 ng/mL

Western Blotting (denaturing and reducing): 0.2 to 2 g/mL. M-1744-50 antibody detects 14 kDa mature BDNF monomer in human serum human and acid treated human and rodent whole tissue homogenates. In simple Tris-homogenate human brain lysates, M-1744-50 detects mature BDNF at 14kDa as well as an uncharacterized band at ~50kDa. No proBDNF band is detected in our samples, although M-1744-50 will detect proBDNF in purified form on blots. Additionally, M-1744-50 detects an additional band 18 kDa BDNF isoform human serum. For cell lysates, some caution is advised as in our hands M-1744-100 fails to detect mature BDNF on SH-SY5Y or C6 cell lysates but does detect mature BDNF in neuronal cell culture lysates. The reason for these differences has not been characterized. For detection of mature BDNF in cell lysates such as SH-SY5Y we recommend affinity-purified rabbit polyclonal antibody to rhBDNF (R-1707-100), or rabbit polyclonal antibody to BDNF peptide 1-10 (R-083-100, whole serum; R-066-500, IgG).

Immunofluorescence: 1 to 4 g/mL, 4% formaldehyde fixation and permeabilization. Staining can be weak until optimized.

M-1744-50 is not recommended for Flow Cytometry at this time.

Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
Specificity Detects human, mouse, rat, guinea pig BDNF. Expected to detect BDNF from other species due to sequence homology.
Blast URL Click here
Form Lyophilized from a solution containing PBS pH 7.4, 3% trehalose, with 0.1% sodium azide.
Reconstitution Spin vial briefly before opening. Reconstitute with 50 L sterile water to obtain a concentration of 1 mg/mL.
Storage Store lyophilized antibody at -20C to -80C protected from moisture. After reconstitution divide antibody into useful aliquots and keep aliquots at -20C to -80C for a higher stability. Working aliquots can be kept at 2-8C for up to 1 month. Avoid repetitive freeze/thaw cycles.
Expiry Date 12 months after purchase if unopened.
References Instructions for Acid-Extraction of Brain Tissue

BDNF Antibodies for Western Blotting
Images (click to zoom)
Mouse monoclonal antibody to rh BDNF [4C8]: IgG Western blot analysis of BDNF expression in acid-treated rodent and human brain samples. Brain samples were acid-treated at pH 4 (refer to References for a detailed extraction method protocol). M-1744-50/100 (1 µg/mL) detects BDNF monomer at 14 kDa. Additional higher MW bands are not characterized, but may be related to the cross-reaction of the secondary anti-mouse-HRP antibody with endogenous, reduced IgG present in samples, in particular mouse tissue.

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 (1/6000), 2 hours at RT; Detection: Chemiluminiscence.
Mouse monoclonal antibody to rh BDNF [4C8]: IgG Western blot analysis of BDNF expression in HSA- and IgG-depleted human serum. Monoclonal BDNF antibody M-1744-50/100 (1 g/mL) detects monomeric BDNF at 14 kDa, and a BDNF isoform at 18 kDa, which is also detected by the polyclonal rabbit BDNF antibody R-084-100. The presence of the BDNF-IR band at 18 kDa in human serum ( Tongiorgi et al., 2012) and mouse hippocampal lysates ( Simmons et al., 2009) has been published. Non-specific higher MW bands are visible for M-1744-50/100, which are due to non-specific binding of the secondary anti-mouse-HRP antibody. Western Blotting Method: SDS-PAGE: denaturing and reducing, 12% Bis-Tris gel; Transfer: Tris-Glycine buffer, semi-dry transfer; Membrane: nitrocellulose (0.22 m); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4C; Secondary antibody: anti-mouse-HRP or anti-rabbit-HRP (1/6000), 2 hours at RT; Detection: Chemiluminiscence.
Mouse monoclonal antibody to rh BDNF [4C8]: IgG Immunofluorescence analysis of BDNF expression in rat C6 glioma cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) C6 cells were incubated with BDNF antibody M-1744-50/100 (2 μg/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (5 μg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). BDNF-IR is observed in the perinuclear cytoplasm. Magnification: 100x.
Mouse monoclonal antibody to rh BDNF [4C8]: IgG Western blot analysis of neuronal cell lysates (Lanes 2 and 3) and rhBDNF (Lane 1). M-1744-100 was used at 1 g/mL.Photo courtesy of Indrek Koppel and Tnis Timmusk (Tallinn Technical University, Institute of Gene Technology)
Mouse monoclonal antibody to rh BDNF [4C8]: IgG Western blot analysis of BDNF expression human brain (Tris-homogenate). Monoclonal mouse antibody to rhBDNF, Clone 4C8 (M-1744-50/100, 1 μg/mL detects monomeric BDNF at 14 kDa. Another band at ~50 kDa is observed which has not been characterized. 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 4C; Secondary antibody: anti-mouse-HRP (1/6000), 2 hours at RT; Detection: Chemiluminiscence.
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