Phospho-EEF2 (T56) Recombinant Rabbit Monoclonal Antibody [PSH06-36]
cat.: HA722670
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | PSH06-36 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| 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: 95 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phosphopeptide corresponding to residues surrounding Thr56 of human EEF2. |
| Positive control: | HeLa starved for 24 hours then add 10μM forskolin for 1 hour cell lysate, NIH/3T3 cell lysate, NIH/3T3 starved for 24 hours then add 10μM forskolin for 1 hour cell lysate, PC-12 treated with 20μM forskolin for 1 hour then add 100nM Calyculin A for 30 minutes cell lysate, HepG2 cell lysate, rat liver tissue, HeLa cells starved for 24 hours then add 10μM forskolin for 1 hour, PC-12 cells treated with 20μM forskolin for 1 hour then add 100nM Calyculin A for 30 minutes. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IHC-P IF-Cell |
1:2,000 1:500 1:100 |
| Uniprot #: | SwissProt: P13639 Human | P58252 Mouse | P05197 Rat |
| Alternative names: | EEF 2 Eef2 EF-2 EF2 EF2_HUMAN Elongation factor 2 Eukaryotic translation elongation factor 2 Polypeptidyl tRNA translocase SCA26 |
Images
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Fig1:
Western blot analysis of Phospho-EEF2 (T56) on different lysates with Rabbit anti-Phospho-EEF2 (T56) antibody (HA722670) at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa starved for 24 hours then add 10μM forskolin for 1 hour cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: NIH/3T3 starved for 24 hours then add 10μM forskolin for 1 hour cell lysate Lane 5: PC-12 cell lysate Lane 6: PC-12 treated with 20μM forskolin for 1 hour then add 100nM Calyculin A for 30 minutes cell lysate Lane 7: HepG2 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 95 kDa Observed band size: 95 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 (HA722670) at 1/2,000 dilution was used in 5% NFDM/TBST 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:
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Phospho-EEF2 (T56) antibody (HA722670) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722670) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig3:
Immunocytochemistry analysis of HeLa cells starved for 24 hours then add 10μM forskolin for 1 hour labeling Phospho-EEF2 (T56) with Rabbit anti-Phospho-EEF2 (T56) antibody (HA722670) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-EEF2 (T56) antibody (HA722670) at 1/100 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 (M1305-2, 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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Fig4:
Immunocytochemistry analysis of PC-12 cells treated with 20μM forskolin for 1 hour then add 100nM Calyculin A for 30 minutes labeling Phospho-EEF2 (T56) with Rabbit anti-Phospho-EEF2 (T56) antibody (HA722670) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-EEF2 (T56) antibody (HA722670) at 1/100 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 (M1305-2, 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.