Anti-FGFR1 (phospho Y654) antibody

• WB

Supplier Data

Western blot - Anti-FGFR1 (phospho Y654) antibody (AB59194)

Loading control : Beta Actin

All lanes:

Western blot - Anti-FGFR1 (phospho Y654) antibody (ab59194) at 1/1000 dilution

Lane 1:

293 untreated lysates at 20 µg

Lane 2:

293 treated with 200 ng/mL EGF for 30 minutes at 20 µg

Lane 3:

HeLa untreated lysates at 20 µg

Lane 4:

HeLa treated with 200 ng/mL EGF for 30 minutes at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (H+L) HRP at 1/10000 dilution

Predicted band size: 92 kDa

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• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FGFR1 (phospho Y654) antibody (AB59194)

IHC image of FGFR1 (phospho Y654) staining in Human breast adenocarcinoma 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 ab59194, 1μg/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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-FGFR1 (phospho Y654) antibody (AB59194)

ICC/IF image of ab59194 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab59194, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FGFR1 (phospho Y654) antibody (AB59194)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using FGFR1 (phospho Y654) antibody (ab59194) at 1/200 dilution. Primary antibody was incubated at 4°C overnight. Antigen retrieval was Tris-EDTA,pH8.0. Secondary antibody was diluted at 1/200 and incubation was at room temperature for 30 minutes.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FGFR1 (phospho Y654) antibody (AB59194)

Immunofluorescent analysis of rat lung tissue labeling FGFR1 (phospho Y654) with ab59194 at 1/200 at 4°C overnight. Alexa flour 594 labeled Secondary antibody was diluted at 1 : 2000 (room temperature, 50min).

• WB

Supplier Data

Western blot - Anti-FGFR1 (phospho Y654) antibody (AB59194)

All lanes:

Western blot - Anti-FGFR1 (phospho Y654) antibody (ab59194) at 1/1000 dilution

All lanes:

HeLa cell lysate

Predicted band size: 92 kDa

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• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FGFR1 (phospho Y654) antibody (AB59194)

Immunofluorescent analysis of COS7 cells labeling FGFR1 (phospho-Tyr654) with ab59194 at 1 : 100. The image on the right is blocked with the phosphopeptide prior to imunnoflurescent labeling.

• WB

Unknown

Western blot - Anti-FGFR1 (phospho Y654) antibody (AB59194)

All lanes:

Western blot - Anti-FGFR1 (phospho Y654) antibody (ab59194) at 1/500 dilution

Lane 1:

293 cell extract treated with insulin (0.01U/ml, 15 mins)

Lane 2:

293 cell extract treated with insulin (0.01U/ml, 15 mins) with immunizing phosphopeptide

Predicted band size: 92 kDa

Observed band size: 120 kDa

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