Anti-FGFR1 (phospho Y654) antibody (ab59194)
Key features and details
- Rabbit polyclonal to FGFR1 (phospho Y654)
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Human, African green monkey
- Isotype: IgG
Overview
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Product name
Anti-FGFR1 (phospho Y654) antibody
See all FGFR1 primary antibodies -
Description
Rabbit polyclonal to FGFR1 (phospho Y654) -
Host species
Rabbit -
Specificity
Binds human and mouse FGFR1 only when phosphorylated at tyrosine 654 and rat FGFR1 only when phosphorylated at tyrosine 561. -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF RatHumanAfrican green monkeyIHC-P MouseHumanWB Human
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Immunogen
Synthetic peptide corresponding to Human FGFR1 aa 626-675. Synthetic phosphopeptide (Human) from around the phosphorylation site of tyrosine 654 (DYYPKK)
Database link: P11362 -
Positive control
- WB: 293 and HeLa cell lysates treated with EGF, 293 cell extract treated with insulin. IHC-P: Human breast adenocarcinoma, mouse kidney. ICC/IF: SKNSH cells. COS7 cells. IF: Rat lung.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride
Without Mg+2 and Ca+2 -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
Affinity purified from rabbit antiserum by affinity chromatography using epitope specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-FGFR1 (phospho Y654) antibody (ab59194)All lanes : Anti-FGFR1 (phospho Y654) antibody (ab59194) at 1/1000 dilution
Lane 1 : 293 untreated lysates
Lane 2 : 293 treated with 200 ng/mL EGF for 30 minutes
Lane 3 : HeLa untreated lysates
Lane 4 : HeLa treated with 200 ng/mL EGF for 30 minutes
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (H+L) HRP at 1/10000 dilution
Predicted band size: 92 kDa
Observed band size: 156 kDa why is the actual band size different from the predicted?Loading control: Beta Actin
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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.
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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).
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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. -
Western blot - Anti-FGFR1 (phospho Y654) antibody (ab59194)All lanes : 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 why is the actual band size different from the predicted?
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Western blot - Anti-FGFR1 (phospho Y654) antibody (ab59194)Anti-FGFR1 (phospho Y654) antibody (ab59194) at 1/1000 dilution + HeLa cell lysate
Predicted band size: 92 kDa
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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.
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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.
