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BDNF Recombinant Rabbit Monoclonal Antibody [SJ12-09]

Specification

Quota

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Catalog# ET1606-42

BDNF Recombinant Rabbit Monoclonal Antibody [SJ12-09]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Predicted reactivity

  • Zebrafish

Predicted species support after-sales service
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

BDNF Recombinant Rabbit Monoclonal Antibody [SJ12-09]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within C-terminal human BDNF.

Species Reactivity

Human, Mouse, Rat (Predicted: Zebrafish)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 28 kDa

Positive Control

SH-SY5Y cell lysate, Neuro-2a cell lysate, mouse brain tissue lysate, rat brain tissue lysate, rat hippocampus tissue lysate, mouse hippocampus tissue lysate, Hela.

Conjugation

unconjugated

Clone Number

SJ12-09

RRID

AB_3069737

Product Features

Form

Liquid

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:20,000

  • IF-Cell

  • 1:100-1:200

  • FC

  • 1:1,000

Target

Function

BDNF acts on certain neurons of the central nervous system and the peripheral nervous system, helping to support survival of existing neurons, and encouraging growth and differentiation of new neurons and synapses. In the brain it is active in the hippocampus, cortex, and basal forebrain—areas vital to learning, memory, and higher thinking. BDNF is also expressed in the retina, kidneys, prostate, motor neurons, and skeletal muscle, and is also found in saliva. BDNF itself is important for long-term memory. Although the vast majority of neurons in the mammalian brain are formed prenatally, parts of the adult brain retain the ability to grow new neurons from neural stem cells in a process known as neurogenesis. Neurotrophins are proteins that help to stimulate and control neurogenesis, BDNF being one of the most active. Mice born without the ability to make BDNF have developmental defects in the brain and sensory nervous system, and usually die soon after birth, suggesting that BDNF plays an important role in normal neural development. Other important neurotrophins structurally related to BDNF include NT-3, NT-4, and NGF. BDNF is made in the endoplasmic reticulum and secreted from dense-core vesicles. It binds carboxypeptidase E (CPE), and disruption of this binding has been proposed to cause the loss of sorting BDNF into dense-core vesicles. The phenotype for BDNF knockout mice can be severe, including postnatal lethality. Other traits include sensory neuron losses that affect coordination, balance, hearing, taste, and breathing. Knockout mice also exhibit cerebellar abnormalities and an increase in the number of sympathetic neurons. Certain types of physical exercise have been shown to markedly (threefold) increase BDNF synthesis in the human brain, a phenomenon which is partly responsible for exercise-induced neurogenesis and improvements in cognitive function. Niacin appears to upregulate BDNF and tropomyosin receptor kinase B (TrkB) expression as well.

Background References

1. Adzic, M. et al. 2015. The contribution of hypothalamic neuroendocrine, neuroplastic and neuroinflammatory processes to lipopolysaccharide-induced depressive-like behaviour in female and male rats: Involvement of glucocorticoid receptor and C/EBP-β. Behavioural brain research. 291: 130-139.

2. Xu, Q. et al. 2015. Sigma 1 receptor activation regulates brain-derived neurotrophic factor through NR2A-CaMKIV-TORC1 pathway to rescue the impairment of learning and memory induced by brain ischaemia/reperfusion. Psychopharmacology. 232: 1779-91.

Sequence Similarity

Belongs to the NGF-beta family.

Tissue Specificity

Detected in blood plasma and in saliva (at protein level). Brain. Highly expressed in hippocampus, amygdala, cerebral cortex and cerebellum. Also expressed in heart, lung, skeletal muscle, testis, prostate and placenta.

Post-translational Modification

[BDNF precursor form]: N-glycosylated and glycosulfated, contrary to mature BDNF.; Mature BDNF is produced by proteolytic removal of the propeptide, catalyzed by a FURIN family member. In addition, the precursor form is proteolytically cleaved within the propeptide, but this is not an obligatory intermediate for the production of mature BDNF. Can be converted into mature BDNF by plasmin (PLG).

Subcellular Location

Secreted.

UNIPROT

SwissProt: P23560 Human

SwissProt: P21237 Mouse

SwissProt: P23363 Rat

Synonyms

Abrineurin antibody

ANON2 antibody

BDNF antibody

BDNF_HUMAN antibody

Brain Derived Neurotrophic Factor antibody

Brain-derived neurotrophic factor antibody

BULN2 antibody

MGC34632 antibody

Neurotrophin antibody

Images

  • Western blot analysis of BDNF on different lysates with Rabbit anti-BDNF antibody (<a href="/products/ET1606-42" style="font-weight: bold;text-decoration: underline;">ET1606-42</a>) at 1/5,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate<br />Lane 2: Neuro-2a cell lysate<br />Lane 3: Mouse brain tissue lysate<br />Lane 4: Rat brain tissue lysate<br />Lane 5: Rat hippocampus tissue lysate<br />Lane 6: Mouse hippocampus tissue lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 28 kDa<br />Observed band size: 28 kDa<br /><br />Exposure time: 1 minute 40 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1606-42" style="font-weight: bold;text-decoration: underline;">ET1606-42</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of BDNF on different lysates with Rabbit anti-BDNF antibody (ET1606-42) at 1/5,000 dilution.

    Lane 1: SH-SY5Y cell lysate
    Lane 2: Neuro-2a cell lysate
    Lane 3: Mouse brain tissue lysate
    Lane 4: Rat brain tissue lysate
    Lane 5: Rat hippocampus tissue lysate
    Lane 6: Mouse hippocampus tissue lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 28 kDa
    Observed band size: 28 kDa

    Exposure time: 1 minute 40 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-42) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of BDNF on different lysates with Rabbit anti-BDNF antibody (<a href="/products/ET1606-42" style="font-weight: bold;text-decoration: underline;">ET1606-42</a>) at 1/20,000 dilution.<br /><br />Lane 1: MDA-MB-231-si NT cell lysate<br />Lane 2: MDA-MB-231-si BDNF cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 28 kDa<br />Observed band size: 28 kDa<br /><br />Exposure time: 14 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1606-42" style="font-weight: bold;text-decoration: underline;">ET1606-42</a>) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/100,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of BDNF on different lysates with Rabbit anti-BDNF antibody (ET1606-42) at 1/20,000 dilution.

    Lane 1: MDA-MB-231-si NT cell lysate
    Lane 2: MDA-MB-231-si BDNF cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 28 kDa
    Observed band size: 28 kDa

    Exposure time: 14 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-42) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of Neuro-2a cells labeling BDNF with Rabbit anti-BDNF antibody (<a href="/products/ET1606-42" style="font-weight: bold;text-decoration: underline;">ET1606-42</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BDNF antibody (<a href="/products/ET1606-42" style="font-weight: bold;text-decoration: underline;">ET1606-42</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Neuro-2a cells labeling BDNF with Rabbit anti-BDNF antibody (ET1606-42) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BDNF antibody (ET1606-42) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of Neuro-2a cells labeling BDNF.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1606-42" style="font-weight: bold;text-decoration: underline;">ET1606-42</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Neuro-2a cells labeling BDNF.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1606-42, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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