Anti-FGFR1 alpha 抗体 [M2F12] (ab829)

Reviews (1)Q&A (3)References (6)

製品の概要

  • 製品名
    Anti-FGFR1 alpha antibody [M2F12]
    FGFR1 alpha 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [M2F12] to FGFR1 alpha
  • 由来種
    Mouse
  • 特異性
    Reacts with alpha isoform of FRFR1.
  • アプリケーション
    適用あり: ICC/IF, IHC-P, IHC-Fr, IP, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Recombinant human ectodomain of FGFr1a expressed in E. coli beginning with pro23; antigen contained NH2-terminal gly-ser-pro-gly-ile and COOH-terminal glu-phe sequences.

  • エピトープ
    NH2-terminus of unique NH2-terminal Ig loop of FGFr1. Epitope is within the sequence between glu30 and ala74 of FGFr1a.
  • ポジティブ･コントロール
    • IHC: Rat prostate tissue.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab829 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 10 µg/ml.
IHC-P Use at an assay dependent concentration. PubMed: 19728793
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 120 kDa.

ターゲット情報

  • 関連性
    FGFR1 (fibroblast growth factor receptor 1) is a member of the fibroblast growth factor receptor family and contains an Ig-like domain and a tyrosine kinase domain. This receptor has multiple isoforms and is a Type I membrane protein. FGFR1 is widely expressed, with distinct isoforms expressed in specific tissues. FGFR1 binds fibroblast growth factor and induces mitogenesis and cellular differentiation. Defects in FGFR1 result in Pfeiffer syndrome associated with craniosynostosis. FGFR1 can be modified by phosphorylation and can bind basic/acidic fibroblast factor depending on the receptor isoform. FGFR1 has been shown to interact with N-cadherin and NCAM.
  • 細胞内局在
    Cell Membrane
  • 参照データベース
    see all
  • 別名
    • BFGFR antibody
    • CD331 antibody
    • CEK antibody
    • FGFBR antibody
    • FGFR1 antibody
    • FLG antibody
    • FLT2 antibody
    • FMS like tyrosine kinase 2 antibody
    • HBGFR antibody
    • KAL2 antibody
    • N SAM antibody
    • OGD antibody
    see all

画像

  • Immunocytochemistry/ Immunofluorescence - Anti-FGFR1 alpha antibody [M2F12] (ab829)
    Immunocytochemistry/ Immunofluorescence - Anti-FGFR1 alpha antibody [M2F12] (ab829)
    ICC/IF image of ab829 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab829, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

参考文献

This product has been referenced in:
  • Borroto-Escuela DO  et al. The existence of FGFR1-5-HT1A receptor heterocomplexes in midbrain 5-HT neurons of the rat: relevance for neuroplasticity. J Neurosci 32:6295-303 (2012). Read more (PubMed: 22553035) »
  • Hellingman CA  et al. FIBROBLAST GROWTH FACTOR RECEPTORS IN IN-VITRO AND IN-VIVO CHONDROGENESIS: RELATING TISSUE ENGINEERING USING ADULT MESENCHYMAL STEM CELLS TO EMBRYONIC DEVELOPMENT. Tissue Eng Part A : (2009). IHC-P ; Mouse . Read more (PubMed: 19728793) »
See all 6 Publications for this product

ab829 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

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Western blot abreview for Anti-FGFR1 alpha antibody [M2F12]

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Neuronal cells)
Gel Running Conditions
Non-reduced Denaturing (4-12%)
Loading amount
25 µg
Specification
Neuronal cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Abcam user community

Verified customer

投稿 Dec 19 2016

Question

Customer would like to know if this antibody binds to FGFR4 as well as FGFR1?

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Answer

No - this antibody is specific for FGFR1.

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Question

Concerning the epitope - does ab829 bind on the cell surface or on the inside?

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Answer

Ab829 binds on the cell surface.

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Question

I will try to answer all your questions below, and I also enclose pictures of staining results of frozen sections. Hope to hear from you. 1. Please describe the problem (high background, no staining etc).High background and weak signal from the antibody 2. On what material are you testing the antibody in IHC? Human pancreas tumors, grwon in SCID mice. ï Species? Human and maybe mouse (ingrowth of mouce cells into the tumor) ï Cell line? ï Tissue? Tumors 3. How did you fix the samples? Acetone Ethanol, methanol Acetone: Acetone Paraformaldehyde Other 4. Did you apply antigen retrieval step? No >ï Enzymatic method >ï Heat mediated technique >ï Other 5. How did you block the unspecific binding sites? 3% bovine serum albumine 6. Primary antibody Species: Human, mouse, rat Dilutions: 1:100, 1:1000 Incubation time: over night at 4 degrees. Washes: 3x5 minutes ï Specification (in which species was it raised against)? ï At what dilution(s) have you tested this antibody? ï Incubation time, wash steps (multiple short washes are more effective than fewer longer wash steps)? 7. Secondary antibody ï What secondary antibody are you using? ï Specification (in which species was it raised against)? ï At what dilution(s) have you tested this antibody? ï Incubation, wash steps? ï Do you know whether the problems you are experiencing come from the >secondary? ï What detection method are you using? Secondary: Alexa goat anti-mouse, HCA (highly cross adsorbed); Molecular Probes. Dilutions: 1:1000 Incubation: 1 hour room temp. Wash: 3x10 min I daubt that the problems are due to the secondary as I use these type of secondary antibodies every week. Detection: immunoflourescence 8. Background staining ï Please provide an image of your staining It is included. 9. Which detection system did you use? Belive I have answered this question in panel 7. 10. Did you apply positive and negative controls along with the samples? Please specify. I always try primary alone, and secondary alone. 11. Optimization attempts ï How many times have you tried the IHC? ï Do you obtain the same results every time? ï What steps have you altered? Times with this antibody: 2 times Yes, the same result but at two different concentrations, which means weaker signal/background with a more diluted primary antibody. I have altered the dilutions. My positive control is the double staining of CD31 in both cases. I also include a picture of FGFR-1 and CD31 staining of FGFR-1 from Santa Cruz. Finally, we would like to reassure you that Abcam policy dictates that if a product does not performed as stated on the datasheet; we will offer a full reimbursement or provide a replacement of equivalent value. We are looking forward to hearing from you soon.

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Answer

Thank you for your patience. I have been discussing this complaint with my colleagues. Unfortunately, we do not really know why this antibody did not perform well for you. We haven't had any complaints about this one. In fact, most of our customers for this antibody use it for immunohistochemistry. A couple of things stand out in your protocol: (1) You pretreat the tissue sections with bovine serum albumin to block non-specific binding. Because BSA doesn't contain any immunoglobulin that can occupy Fc receptors that might be present on cells in tissues, we always recommend blocking tissue sections with an unrelated normal serum, such as goat or rabbit. We realize, though, that you feel that blocking with BSA worked well for the Santa Cruz antibody, so you did probably not inclined to try blocking with normal serum. (2) It was difficult to compare the photos of the tissue staining that you provided because each photo appeared to be a different tissue section than the other photos. Where your felt that you saw very high background staining (with Abcam antibody at 1:100), we think you used too high a concentration of primary antibody. According to our records, the batch that you purchased was at a concentration of 1.65 mg/ml. Therefore, a 1:100 dilution of this would give a working solution of 16.5 ug/ml. This is a rather high antibody concentration for IHC. Most of our antibodies work in the range of 3-5 ug/ml. Of course, the 1:1000 dilution gives a working solution of 1.65 ug/ml, and this might be too low. We could send you a replacement vial, but it would be the same lot as what you already received (it is the only lot we have at this time), and we would guess that you wouldn't be eager to try this one again. Perhaps the most expedient solution is for you a credit note or refund. Please do let me know what you would like to do.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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