Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized U937 (Human histiocytic lymphoma cell line) cells labeling Iba1 with ab178847 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on U937 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows:-

-ve control 1: ab178847 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Iba1 with ab178847 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on microglia of the normal Human cerebrum is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

ab33922 showing positive staining in Normal brain tissue.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-N-F1 cells labeling TREM2 with ab209814 at 1/500 dilution, followed by Donkey anti rabbit IgG (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution (green).

The nuclear counter stain is DAPI (blue).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling ICAM1 with ab53013 at 1/1000. Heat mediated antigen retrieval was performed using ab93684, Tris/EDTA buffer pH 9. A ready to use HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody. Counterstained with hematoxylin.

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)

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.

All lanes : Anti-TNF alpha antibody [EPR20972] (ab215188) at 1/1000 dilution

Lane 1 : THP-1 (human monocytic leukemia cell line) differentiated with 100 nM TPA overnight, whole cell lysate

Lane 2 : THP-1 (human monocytic leukemia cell line) differentiated with 100 nM TPA overnight, then treated with 100 ng/ml lipopolysaccharides (LPS) for 7 hours with addition of 1 µg/ml brefeldin A (BFA) for the last 3 hours, whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary

All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Developed using the ECL technique.

Predicted band size: 26 kDa

Observed band size: 26 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.