Anti-NGF antibody [EP1320Y]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-NGF antibody [EP1320Y] (AB52918)

Immunofluorescence staining of U87-MG cells with purified ab52918 at a working dilution of 1 in 300 counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (ab7291) and Alexa Fluor^® 594 goat anti-mouse at a dilution of 1/500 (ab150120) . The secondary antibody was ab150077 Alexa Fluor^® 488 goat anti rabbit used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in the bottom middle and right hand panels - for the first negative control purified ab52918 was used at a dilution of 1/200 followed by an Alexa Fluor^® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab15007) were used.

• WB

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Western blot - Anti-NGF antibody [EP1320Y] (AB52918)

All lanes:

Western blot - Anti-NGF antibody [EP1320Y] (ab52918) at 1/500 dilution

All lanes:

fetal thyroid tissue at 10 µg

Secondary

All lanes:

Goat anti rabbit IgG HRP antibody at 1/2000 dilution

Predicted band size: 27 kDa

Observed band size: 27 kDa

false

• WB

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Western blot - Anti-NGF antibody [EP1320Y] (AB52918)

Blocking Buffer : 5% NFDM/TBST

Dilution Buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-NGF antibody [EP1320Y] (ab52918) at 1/1000 dilution

All lanes:

Mouse thyroid lysate at 20 µg

Secondary

All lanes:

HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 32 kDa

false

• WB

Unknown

Western blot - Anti-NGF antibody [EP1320Y] (AB52918)

Blocking Buffer : 5% NFDM/TBST

Dilution Buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-NGF antibody [EP1320Y] (ab52918) at 1/2000 dilution

All lanes:

Rat thyroid lysate at 20 µg

Secondary

All lanes:

HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 32 kDa

false

• WB

Unknown

Western blot - Anti-NGF antibody [EP1320Y] (AB52918)

Blocking Buffer : 5% NFDM/TBST

Dilution Buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-NGF antibody [EP1320Y] (ab52918) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 20 µg

Lane 2:

Human fetal thymus lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 27 kDa

Observed band size: 32 kDa

false

• WB

Supplier Data

Western blot - Anti-NGF antibody [EP1320Y] (AB52918)

The positive cell lines (Lanes 7-9) are consistent with the literature : PMID : 12944907, Fig1.

Blocking buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-NGF antibody [EP1320Y] (ab52918) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 20 µg

Lane 2:

Rat brain lysate at 20 µg

Lane 3:

Human cerebellum lysate at 20 µg

Lane 4:

Mouse cerebellum lysate at 20 µg

Lane 5:

Rat cerebellum lysate at 20 µg

Lane 6:

Hela (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) 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

Predicted band size: 27 kDa

Observed band size: 32 kDa

true

Exposure time: 20s