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39788
Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free)
Primary Antibodies
Monoclonal Antibody

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free) #39788

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  1. WB
  2. IHC
  3. IF
  4. F
Western Blotting Image 1: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free)
Western blot analysis of extracts from 293T cells using 4E-BP1 Antibody #9452 (upper) and Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb #2855 (lower). The cells were starved for 24 hours in serum-free medium and underwent a 1 hour amino acid deprivation. Amino acids were replenished for 1 hour. Cells were then either untreated (-) or treated with 100 nM insulin (+) for 30 minutes. Data were generated using the standard formulation of this product.
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Immunohistochemistry Image 1: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb in the presence of control peptide (left) or Phospho-4E-BP1 (Thr37/46) Blocking Peptide #1052 (right). Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 3: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human lymphoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 4: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right), using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101. Data were generated using the standard formulation of this product.
Immunofluorescence Image 1: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free)
Confocal immunofluorescent analysis of 293 cells, expressing either non-targeting shRNA (top) or shRNA targeting 4E-BP1/2 (bottom), using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (green). To confirm phospho-specificity, cells were treated with an inhibitor cocktail consisting of LY294002 #9901, U0126 #9903, and Rapamycin #9904 (50 μM, 10 μm, 10 nM, 2 hr; left), stimulated with insulin (100 nM, 30 min; middle), or processed with λ-phosphatase following insulin stimulation (right). Red = Propidium Iodide (PI)/RNase Staining Solution #4087. Data were generated using the standard formulation of this product.
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Flow Cytometry Image 1: Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free)
Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, Wortmannin #9951, and U0126 #9903 (50 μM, 1 μM, and 10 μM, 2 hr; blue), using Phospho-4E-BP1 (Thr36/46) (236B4) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Data were generated using the standard formulation of this product.
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To Purchase # 39788
製品# サイズ 数量 価格 在庫
39788SF
100 µg

Supporting Data

REACTIVITY H M R Mk Dm
SENSITIVITY
MW (kDa) 15 to 20
Source/Isotype Rabbit IgG

Application Key:

  • WB-Western Blot
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • C&R-CUT&RUN
  • C&T-CUT&Tag
  • DB-Dot Blot
  • eCLIP-eCLIP
  • IF-Immunofluorescence
  • F-Flow Cytometry

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Vir-Virus
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Usage Information

This product is the carrier free version of product #2855. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol. This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN, or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

Formulation

Supplied in 1X PBS, BSA and Azide Free.

For standard formulation of this product see product #2855.

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity / Sensitivity

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (BSA and Azide Free) detects endogenous levels of 4E-BP1 only when phosphorylated at Thr37 and/or Thr46. This antibody may cross-react with 4E-BP2 and 4E-BP3 when phosphorylated at equivalent sites. Non-specific staining has been observed in mitotic cells by immunofluorescence.

Species Reactivity:

Human, Mouse, Rat, Monkey, D. melanogaster

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr37 and Thr46 of mouse 4E-BP1.

Background

Translation repressor protein 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation by binding to the translation initiation factor eIF4E. Hyperphosphorylation of 4E-BP1 disrupts this interaction and results in activation of cap-dependent translation (1). Both the PI3 kinase/Akt pathway and FRAP/mTOR kinase regulate 4E-BP1 activity (2,3). Multiple 4E-BP1 residues are phosphorylated in vivo (4). While phosphorylation by FRAP/mTOR at Thr37 and Thr46 does not prevent the binding of 4E-BP1 to eIF4E, it is thought to prime 4E-BP1 for subsequent phosphorylation at Ser65 and Thr70 (5).
  1. Pause, A. et al. (1994) Nature 371, 762-7.
  2. Brunn, G.J. et al. (1997) Science 277, 99-101.
  3. Gingras, A.C. et al. (1998) Genes Dev 12, 502-13.
  4. Fadden, P. et al. (1997) J Biol Chem 272, 10240-7.
  5. Gingras, A.C. et al. (1999) Genes Dev 13, 1422-37.

Pathways & Proteins

Explore pathways + proteins related to this product.

使用に関する制限

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For Research Use Only. Not For Use In Diagnostic Procedures.
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