BDNF Recombinant Rabbit Monoclonal Antibody [SJ12-09]
cat.: HA750104
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 2-8℃. Avoid freeze.
Storage buffer: 1*PBS (pH7.4).
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: U-87 MG (Human glioblastoma cell) cell lysate, HeLa (Human cervical adenocarcinoma cell) cell lysate, C6 (Rat glioma cell) cell lysate, Mouse brain tissue lysate, Mouse cerebellum tissue lysate, Rat brain tissue lysate, Rat cerebellum tissue lysate, MDA-MB-231 cell lysate, Neuro-2a.
Subcellular location: Secreted.
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
HA750104_1.jpg Fig1: This data was developed using HA750104, the same antibody clone in a different buffer formulation.
Western blot analysis of BDNF on different lysates with Rabbit anti-BDNF antibody (HA750104) at 1/5,000 dilution.

Lane 1: U-87 MG (Human glioblastoma cell) cell lysate (15 µg/Lane)
Lane 2: HeLa (Human cervical adenocarcinoma cell) cell lysate (15 µg/Lane)
Lane 3: C6 (Rat glioma cell) cell lysate (15 µg/Lane)
Lane 4: Mouse brain tissue lysate (30 µg/Lane)
Lane 5: Mouse cerebellum tissue lysate (30 µg/Lane)
Lane 6: Rat brain tissue lysate (30 µg/Lane)
Lane 7: Rat cerebellum tissue lysate (30 µg/Lane)

Exposure time: 30 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA750104, 1/5,000 in 5% NFDM/TBST, overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 28 kDa
Observed band size: 25-40 kDa
HA750104_2.jpg Fig2: This data was developed using HA750104, the same antibody clone in a different buffer formulation.
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.
HA750104_3.jpg Fig3: This data was developed using HA750104, the same antibody clone in a different buffer formulation.
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.
HA750104_4.jpg Fig4: This data was developed using HA750104, the same antibody clone in a different buffer formulation.
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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.