Anti-FGFR1 antibody [EPR806Y] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-FGFR1 antibody [EPR806Y] - BSA and Azide free (AB271861)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling FGFR1 with at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in SH-SY5Y cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76464).

• IP

Lab

Immunoprecipitation - Anti-FGFR1 antibody [EPR806Y] - BSA and Azide free (AB271861)

ab76464 (purified) at 1/190 immunoprecipitating FGFR1 in 10 μg A-204 (Human muscle rhabdomyosarcoma)whole cell lysate (Lanes 1 and 2, observed at 145 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730) instead of ab76464 in A-204 whole cell lysate. For western blotting, ab76464 at 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Blocking/Dilution buffer and concentration : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76464).

All lanes:

Immunoprecipitation - Anti-FGFR1 antibody [EPR806Y] (<a href='/en-us/products/primary-antibodies/fgfr1-antibody-epr806y-ab76464'>ab76464</a>)

Predicted band size: 92 kDa

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• WB

Lab

Western blot - Anti-FGFR1 antibody [EPR806Y] - BSA and Azide free (AB271861)

This data was developed using the same antibody clone in a different buffer formulation (ab76464).

Western blot : Anti-FGFR1 antibody [EPR806Y] (ab76464) staining at 1/500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab76464 was shown to bind specifically to FGFR1. A band was observed at 92 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in FGFR1 knockout cell line. To generate this image, wild-type and FGFR1 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-FGFR1 antibody [EPR806Y] (<a href='/en-us/products/primary-antibodies/fgfr1-antibody-epr806y-ab76464'>ab76464</a>) at 1/500 dilution

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

FGFR1 knockout U-87 MG cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 92 kDa

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