Revision 1

#48267Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
mGluR1 (D5H10) Rabbit mAb 12551 20 µl 145, >300 kDa Rabbit IgG
mGluR5 (D6E7B) Rabbit mAb 55920 20 µl 150, 300 kDa Rabbit IgG
FMRP (D14F4) Rabbit mAb 7104 20 µl 80 kDa Rabbit IgG
FXR1 (D10A2) XP® Rabbit mAb 12295 20 µl 78-80, 82-84 kDa Rabbit IgG
FXR2 (D85D6) Rabbit mAb 7098 20 µl 95 kDa Rabbit IgG
CYFIP1 Antibody 44353 20 µl 145 kDa Rabbit 
Phospho-eEF2 (Thr56) Antibody 2331 20 µl 95 kDa Rabbit 
eEF2 Antibody 2332 20 µl 95 kDa Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Fragile X/FMRP Signaling Pathway Antibody Sampler Kit provides an economical means of detecting signaling components of the Fragile X/FMRP signaling pathway. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

Fragile X syndrome, a frequent cause of inherited mental retardation, often results from expansion of the CGG trinucleotide repeat in the gene that encodes the fragile X mental retardation protein (FMRP, [1]). FMRP (also known as FMR1) and its two autosomal homologs (FXR1 and FXR2) all bind RNA and play a role in the pathogenesis of fragile X syndrome (1-3). Each of these related proteins can associate with one another as well as form homodimers and complexes with other RNA-binding proteins like cytoplasmic FMRP interacting protein 1 (CYFIP1, [3,4]). FMRP, FXR1, FXR2, and CYFIP1 have been implicated in the translational regulation of mRNAs (5,6). Importantly, this complex of proteins may be dynamically regulated to drive protein synthesis-dependent forms of synaptic plasticity in response to specific activity. That is, activation of metabotropic glutamate receptors, including mGluR1 and mGlur5, can regulate FMRP-dependent forms of translation via post-translational modification of eukaryotic elongation factor 2 (eEF2) to locally control dynamic translation of important synaptic proteins, which, subsequently, alter synaptic function (7-9).   

  1. Verkerk, A.J. et al. (1991) Cell 65, 905-14.
  2. Siomi, M.C. et al. (1995) EMBO J 14, 2401-8.
  3. Zhang, Y. et al. (1995) EMBO J 14, 5358-66.
  4. Abekhoukh, S. et al. (2017) Dis Model Mech 10, 463-74.
  5. Linder, B. et al. (2008) Hum Mol Genet 17, 3236-46.
  6. De Rubeis, S. et al. (2013) Neuron 79, 1169-82.
  7. Park, S. et al. (2008) Neuron 59, 70-83.
  8. Barnes, S.A. et al. (2015) J Neurosci 35, 15073-81.
  9. Paul, A. et al. (2019) Front Mol Neurosci 12, 97.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    Revision 1
    #48267

    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit

    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts from various cell lines using FXR1 (D10A2) XP® Rabbit mAb.
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from neonatal mouse brain and rat brain using mGluR1 (D5H10) Rabbit mAb.
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from C6 cells, untreated or forskolin-treated (10 µM for 60 minutes), using Phospho-eEF2 (Thr56) Antibody (upper) or eEF2 Antibody #2332 (lower).
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from C6 cells, untreated or forskolin-treated (10 µM for 60 minutes), using Phospho-eEF2 (Thr56) Antibody (upper) or eEF2 Antibody #2332 (lower).
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of CAKI-1 and SU-DHL-4 cells, and human brain extracts, using CYFIP1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from the indicated tissues using mGluR5 (D6E7B) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 7 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from various cell lines using FXR2 (D85D6) Rabbit mAb.
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell lines using FMRP (D14F4) Rabbit mAb.
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from human and mouse skeletal muscle tissues using FXR1 (D10A2) XP® Rabbit mAb.
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse cerebellum using mGlur1 (D5H10) Rabbit mAb.
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 12 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using eEF2 Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of mGluR5 from mouse brain extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is mGluR5 (D6E7B) Rabbit mAb. Western blot analysis was performed using mGluR5 (D6E7B) Rabbit mAb.
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of C2C12 cells, untreated (left) or MG132-treated (10 μg/mL, 3 hr, right) using FXR2 (D85D6) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 15 Expand Image
    Confocal immunofluorescent analysis of C2C12 cells, untreated (left) or treated with MG132 (10 μg/mL, 3 hrs; right), using FMRP (D14F4) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 16 Expand Image
    Confocal immunofluorescent analysis of C2C12 cells, untreated (left) or MG-132 treated (10 μg/mL, 3 hr; right), using FXR1 (D10A2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded rat retina using mGlur1 (D5H10) Rabbit mAb.
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 18 Expand Image
    Confocal immunofluorescent analysis of mouse hippocampus (left) and striatum (right) using mGluR5 (D6E7B) Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Fragile X/FMRP Signaling Pathway Antibody Sampler Kit: Image 19 Expand Image
    Confocal immunofluorescent analysis of adult mouse cerebellum tissue using mGluR1 (D5H10) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).