BDNF Recombinant Rabbit Monoclonal Antibody [SJ12-09]
cat.: ET1606-42
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Zebrafish
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: SJ12-09
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 28 kDa
Isotype: IgG
Immunogen: Synthetic peptide within C-terminal human BDNF.
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.
Subcellular location: Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  FC

1:2,000-1:20,000
1:50-1:200
1:1,000
Uniprot #: SwissProt: P23560 Human | P21237 Mouse | P23363 Rat
Alternative names: Abrineurin ANON2 BDNF BDNF_HUMAN Brain Derived Neurotrophic Factor Brain-derived neurotrophic factor BULN2 MGC34632 Neurotrophin
Images
ET1606-42_1.jpg Fig1: Western blot analysis of BDNF on different lysates with Rabbit anti-BDNF antibody (ET1606-42) at 1/2,000 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 1/2,000 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.
ET1606-42_2.jpg Fig2: 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.
ET1606-42_3.jpg Fig3: ICC staining of BDNF in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1606-42, 1/200) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1606-42_4.jpg Fig4: 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.
ET1606-42_5.jpg Fig5: 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.