Phospho-eIF4EBP1 (S65) Recombinant Rabbit Monoclonal Antibody [PSH09-44]
cat.: HA723106
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH09-44 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 13 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser65 of human eIF4EBP1. |
| Positive control: | 293T cells starved for 24 hours then treated with 100nM insulin for 30 minutes. |
| Subcellular location: | Cytoplasm, cytosol. |
| Recommended Dilutions:
WB IF-Cell IP |
1:1,000 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: Q13541 Human |
| Alternative names: | 4E-BP1 4EBP1 4EBP1_HUMAN BP 1 eIF4E binding protein 1 eIF4E-binding protein 1 Eif4ebp1 Eukaryotic translation initiation factor 4E-binding protein 1 PHAS-I PHASI Phosphorylated heat- and acid-stable protein regulated by insulin 1 |
Images
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Fig1:
Western blot analysis of Phospho-eIF4EBP1 (S65) on different lysates with Rabbit anti-Phospho-eIF4EBP1 (S65) antibody (HA723106) at 1/1,000 dilution. Lane 1: 293T starved for 24 hours cell lysate Lane 2: 293T starved for 24 hours then treated with 100nM insulin for 30 minutes cell lysate Lane 3: 293T starved for 24 hours then treated with 100nM insulin for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 13 kDa Observed band size: 20 kDa Exposure time: 3 minutes; ECL: K1802; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723106) at 1/1,000 dilution was used in primary antibody dilution (WB500D) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of 293T cells starved for 24 hours then treated with 100nM insulin for 30 minutes labeling Phospho-eIF4EBP1 (S65) with Rabbit anti-Phospho-eIF4EBP1 (S65) antibody (HA723106) at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-eIF4EBP1 (S65) antibody (HA723106) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Phospho-eIF4EBP1 (S65) was immunoprecipitated from 0.2 mg 293T cell lysate with HA723106 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723106 at 1/1,000 dilution. Alpaca anti-Rabbit IgG Fc secondary antibody (HA1031) at 1/50,000 dilution was used for 1 hour at room temperature. Lane 1: 293T cell lysate (input) Lane 2: HA723106 IP in 293T cell lysate Lane 3: Rabbit IgG instead of HA723106 in 293T cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 18 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.