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BDNF Recombinant Rabbit Monoclonal Antibody [SJ12-09] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750104

BDNF Recombinant Rabbit Monoclonal Antibody [SJ12-09] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Predicted reactivity

  • Zebrafish

Conjugation

  • unconjugated

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

Overview

Product Name

BDNF Recombinant Rabbit Monoclonal Antibody [SJ12-09] - BSA and Azide free

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

Target 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

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

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

Brain-derived neurotrophic factors (Bdnf), or abrineurin, is a protein that, in humans, is encoded by the BDNF gene. BDNF is a member of the neurotrophin family of growth factors, which are related to the canonical nerve growth factor (NGF), a family which also includes NT-3 and NT-4/NT-5. Neurotrophic factors are found in the brain and the periphery. BDNF was first isolated from a pig brain in 1982 by Yves-Alain Barde and Hans Thoenen. BDNF activates the TrkB tyrosine kinase receptor. BDNF acts on certain neurons of the central nervous system and the peripheral nervous system expressing TrkB, 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.

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/HA750104" style="font-weight: bold;text-decoration: underline;">HA750104</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/HA750104" style="font-weight: bold;text-decoration: underline;">HA750104</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 (HA750104) 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 (HA750104) 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/HA750104" style="font-weight: bold;text-decoration: underline;">HA750104</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/HA750104" style="font-weight: bold;text-decoration: underline;">HA750104</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 (HA750104) 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 (HA750104) 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/HA750104" style="font-weight: bold;text-decoration: underline;">HA750104</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/HA750104" style="font-weight: bold;text-decoration: underline;">HA750104</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 (HA750104) 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 (HA750104) 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/HA750104" style="font-weight: bold;text-decoration: underline;">HA750104</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 (HA750104, 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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