Anti-BDNF antibody [3C11]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BDNF antibody [3C11] (AB203573)

IHC image of BDNF staining in Normal Human Hippocampus formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab203573, 5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BDNF antibody [3C11] (AB203573)

Immunofluorescent analysis of proBDNF expressing U2OS cells labeling BDNF with ab203573 at 10 μg/mL (Left hand panel). Goat anti-mouse AlexaFluor488 was used as secondary antibody. For nuclear staining DAPI was used. Right hand panel : Negative control (non-transfected U2OS cells).

• WB

Lab

Western blot - Anti-BDNF antibody [3C11] (AB203573)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab203573 (1/1000) overnight at 4°C. Antibody binding was detected using goat anti-mouse IgG IR-680 (green) at a 1 : 10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx

All lanes:

Western blot - Anti-BDNF antibody [3C11] (ab203573) at 5 µg/mL

Lane 1:

Human hippocampus at 20 µg

Lane 2:

Rat hippocampus at 20 µg

Lane 3:

Mouse hippocampus at 20 µg

Secondary

All lanes:

goat anti-mouse IgG IR-680 (green) at 1/10000 dilution

Predicted band size: 27 kDa

Observed band size: 15 kDa,40 kDa

false

• WB

Supplier Data

Western blot - Anti-BDNF antibody [3C11] (AB203573)

All lanes:

Western blot - Anti-BDNF antibody [3C11] (ab203573) at 5 µg/mL

All lanes:

CHO supernatant containing BDNF at 10 µL

Secondary

All lanes:

HRP-conjugated goat anti-mouse

Predicted band size: 27 kDa

false

• WB

CiteAb

Western blot - Anti-BDNF antibody [3C11] (AB203573)

BDNF western blot using anti-BDNF antibody [3C11] ab203573. Publication image and figure legend from Zalewska, T., Jaworska, J., et al., 2020, Int J Mol Sci, PubMed 32471267.

ab203573 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab203573 please see the product overview.

Effect of TSA on the expression of BDNF protein in the brain hemispheres after HI. (A) Representative immunoblots of BDNF protein level. The intensity of each band was quantified and normalized in relation to actin. (B) The bar graphs represent the statistical analysis of immunoreactive bands estimated in indicated experimental groups : sham-control (Sham), TSA-treated sham-control (Sham+TSA), hypoxia-ischemia (HI), TSA-treated hypoxia-ischemia (HI+TSA). Note that TSA application increases the level of BDNF protein at 7 days of recovery. The values are mean ± SD from 5 animals per group and time point assessed in 3 independent experiments. Two-way ANOVA tests indicate significant differences between investigated groups, ** p < 0.01, ++++p < 0.0001 (effect of TSA treatment). Abbreviations : ipsi—ipsilateral, contra—contralateral.

false