Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling BDNF with Purified ab108319 at 1 : 500 (0.6 μg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488 ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab108319).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

This data was developed using the same antibody clone in a different buffer formulation (ab108319).

Immunohistochemical analysis of formalin fixed paraffin embedded human brain (cerebrum) labelling BDNF with ab108319 at a concentration of 0.56 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab108319 Anti-BDNF antibody [EPR1292] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling BDNF with purified ab108319 at 1/30 dilution (10 μg/ml) (red). Cells were fixed with 80% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108319).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

This IHC data was generated using the same anti-BDNF antibody clone EPR1292 in a different buffer formulation (cat# ab108319).

Immunohistochemical analysis of paraffin-embedded Human brain tissue using ab108319 at 1/100

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder cancer tissue sections labeling BDNF with Purified ab108319 at 1 : 500 dilution (0.56 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer pH 9.0)

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab108319).

• ICC/IF

AbReview56343****

Immunocytochemistry/ Immunofluorescence - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

This ICC/IF data was generated using the same anti-BDNF antibody clone, EPR1292, in a different buffer formulation (cat# ab108319).

Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling BDNF with ab108319 at a dilution of 1/750. Cells were fixed with Paraformaldehyde and permeabilised with 0.5% Triton-X100 in PBS. ab150081 (1/200) was used as the secondary antibody.

The antibody produces a strong, golgi-associated labelling pattern in both PF and MeOH fixed samples.

This image is courtesy of an Abreview submitted by Kirk McManus (1894904)

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

Immunohistochemistry (Frozen sections) analysis of rat cerebral cortex tissue sections labeling BDNF with Purified ab108319 at 1/100 (2.8 μg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. DAPI was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108319).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

Immunohistochemistry (Frozen sections) analysis of mouse cerebrum tissue sections labeling BDNF with Purified ab108319 at 1/100 (2.8 μg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. DAPI was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108319).

• WB

Lab

Western blot - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

This data was developed using ab108319, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-BDNF antibody [EPR1292] (<a href='/en-us/products/primary-antibodies/bdnf-antibody-epr1292-ab108319'>ab108319</a>) at 1/1000 dilution

Lane 1:

Human brain lysates at 20 µg

Lane 2:

Mouse brain lysates at 20 µg

Lane 3:

Rat brain lysates at 20 µg

Lane 4:

Human hippocampus lysates at 20 µg

Lane 5:

Mouse hippocampus lysates at 20 µg

Lane 6:

Rat hippocampus lysates at 20 µg

Lane 7:

Human cerebellum lysates at 20 µg

Lane 8:

Mouse cerebellum lysates at 20 µg

Lane 9:

Rat cerebellum lysates at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 27 kDa

Observed band size: 15-45 kDa

false

• WB

Lab

Western blot - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108319).

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 unpurified ab108319 (1/1000) overnight at 4°C. ab8245 (mouse anti-GAPDH; 0.05 ug/mL) was included as a loading control. Antibody binding was detected using goat anti-rabbit IgG IR-680 (green) and goat anti-mouse IgG IR800 (red) 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 [EPR1292] (<a href='/en-us/products/primary-antibodies/bdnf-antibody-epr1292-ab108319'>ab108319</a>) at 1/1000 dilution

Lane 1:

Human hippocampus lysate at 20 µg

Lane 2:

Rat hippocampus lysate at 20 µg

Lane 3:

Mouse hippocampus lysate at 20 µg

Secondary

All lanes:

Gt anti Rb IR680 at 1/10000 dilution

Predicted band size: 27 kDa

Observed band size: 15 kDa,28 kDa,35 kDa,45 kDa

false

• WB

Lab

Western blot - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

This data was developed using ab108319, the same antibody clone in a different buffer formulation. Different batches of ab108319 were tested on Mouse brain lysate at 0.3 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 14-45 kDa.

All lanes:

Western blot - Anti-BDNF antibody [EPR1292] (<a href='/en-us/products/primary-antibodies/bdnf-antibody-epr1292-ab108319'>ab108319</a>)

Predicted band size: 27 kDa

false

• WB

Lab

Western blot - Anti-BDNF antibody [EPR1292] - Low endotoxin, Azide free (AB216443)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108319).

Blocking and diluting buffer and concentration : 5% NFDM/TBST

ab213204 was used as a His tag loading control.

All lanes:

Western blot - Anti-BDNF antibody [EPR1292] (<a href='/en-us/products/primary-antibodies/bdnf-antibody-epr1292-ab108319'>ab108319</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Human BDNF expression vector containing a His-tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a Human NGF expression vector containing a His-tag whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a Human NTF3 expression vector containing a His-tag whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a Human NTF4 expression vector containing a His-tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35 kDa

false

Exposure time: 1s