FUNCTION: SwissProt: P68104 # This protein promotes the GTP-dependent binding of aminoacyl-tRNA to the A-site of ribosomes during protein biosynthesis.
SIZE: 462 amino acids; 50141 Da
SUBUNIT: Found in a nuclear export complex with XPO5, EEF1A1, Ran and aminoacylated tRNA. Interacts with XPO5.
SUBCELLULAR LOCATION: Cytoplasm.
TISSUE SPECIFICITY: Brain, placenta, lung, liver, kidney, pancreas but barely detectable in heart and skeletal muscle.
SIMILARITY: SwissProt: P68104 ## Belongs to the GTP-binding elongation factor family. EF-Tu/EF-1A subfamily.

Crude calmodulin-binding proteins from Trypanosoma brucei isolated using calmodulin-affinity chromatography.

EF1α (Elongation factor-1 alpha)

Immunoprecipitation from RIP Lysate:
Representative lot data.
RIP lysate from HeLa cells (2 X 10⁶ cell equivalents per IP) was subjected to immunoprecipitation using 0.5 µg of either a normal mouse IgG or Anti-EF1α antibody. Precipitated proteins (Lane 1: mouse IgG, Lane 2: anti-EF1α) and HeLa whole cell lysate (Lane 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-EF1α antibody (1.0 µg/mL).
Proteins were visualized using One-Step IP-Western kit (GenScript Cat. # L00232) (please see figures).

Automated Microfluidics-based Electrophoretic RNA Separation and Analysis:
RIP Lysate prepared from HeLa cells (2 X 10⁷ cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG or Anti-EF1α antibody and the TM=["MAGNA RIP"] RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of EF1a-associated RNA was verified by automated microfluidics-based electrophoretic RNA separation and analysis. Please refer to the TM=["MAGNA RIP"] (Cat. # 17-700) or EZ-TM=["MAGNA RIP"] (Cat. # 17-701) protocol for experimental details.

Electropherograms were generated by plotting fluorescence intensities versus migration times.
(Please see figures)

The virtual gel view was created from this data.
(Please see figures)

10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).

Fluorescence-based Quantifiation Assay:
Successful immunoprecipitation of EF1α-associated RNA was verified by enrichment over mock immunoprecipitation using a fluorescence-based quantification assay.
Please refer to the Magna RIP (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

53 kDa

Anti-EF1α (Mouse Monoclonal IgG1κ). One vial containing 50 μg of protein G purified mouse monoclonal IgG1κ in 72 μL of 70% storage buffer containing (0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide) and 30% glycerol. Store at -20°C.

Normal Mouse IgG.
One vial containing 125 μg of purified mouse IgG in 125 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.

Stable for 1 year at -20°C from date of receipt.Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Human. Predicted to react with most mammals, and yeast.

Includes negative control mouse IgG antibody.

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