eEF1A1 Recombinant Rabbit Monoclonal Antibody [JB44-13]
Catalog# ET7107-75
eEF1A1 Recombinant Rabbit Monoclonal Antibody [JB44-13]
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WB
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IHC-P
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FC
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IF-Cell
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IF-Tissue
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
eEF1A1 Recombinant Rabbit Monoclonal Antibody [JB44-13]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human eEF1A1 aa 413-462 / 462.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, FC, IF-Cell, IF-Tissue
Molecular Weight
Predicted band size: 50 kDa
Positive Control
HeLa cell lysate, MCF7 cell lysate, 293T cell lysate, Jurkat cell lysate, HepG2 cell lysate, A431 cell lysate, THP-1 cell lysate, K-562 cell lysate, C2C12 cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, C6 cell lysate, Rat kidney tissue lysate, human breast cancer tissue, human kidney tissue, mouse kidney tissue, rat kidney tissue, HeLa, C2C12, PC-12.
Conjugation
unconjugated
Clone Number
JB44-13
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IF-Cell
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1:100-1:250
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IF-Tissue
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1:50-1:200
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IHC-P
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1:200-1:1,000
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FC
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1:1,000
Target
Function
The elongation factor-1 complex is composed of two subunits, EF-1 α1 (elongation factor 1-alpha 1) and EF-1 α2 (elongation factor 1-alpha 2), and is responsible for the delivery of aminoacyl tRNAs to the ribosome. EF-1 α1 is expressed predominately in brain, placenta, lung, liver, kidney and pancreas, while EF-1 α2 is highly expressed in heart, brain and skeletal muscle. Both EF-1 α1 and α2 localize to the nucleus and belong to the GTP-binding elongation factor family. The gene encoding EF-1 α2, which maps to human chromosome 20q13.3, may play a role in the development of ovarian cancer, while the EF-1 α1 gene, mapping to chromosome 6Q14.1, is commonly present as an autoantigen in patients with Felty syndrome. Felty syndrome is a disorder characterized by rheumatoid arthritis, a swollen spleen, decreased white blood cell count, and increased susceptibility to infection.
Background References
1. Maruyama T et al. Txk, a member of the non-receptor tyrosine kinase of the Tec family, forms a complex with poly(ADP-ribose) polymerase 1 and elongation factor 1alpha and regulates interferon-gamma gene transcription in Th1 cells. Clin Exp Immunol 147:164-175 (2007).
2. Boratko A et al. Elongation factor-1A1 is a novel substrate of the protein phosphatase 1-TIMAP complex. Int J Biochem Cell Biol 69:105-113 (2015).
Sequence Similarity
Belongs to the TRAFAC class translation factor GTPase superfamily. Classic translation factor GTPase family. EF-Tu/EF-1A subfamily.
Tissue Specificity
Brain, placenta, lung, liver, kidney, pancreas but barely detectable in heart and skeletal muscle.
Post-translational Modification
ISGylated.; Phosphorylated by TXK. Phosphorylation by PASK increases translation efficiency. Phosphorylated by ROCK2.; Trimethylated at Lys-79 by EEF1AKMT1. Methylated at Lys-165 by EEF1AKMT3, methylation by EEF1AKMT3 is dynamic as well as inducible by stress conditions, such as ER-stress, and plays a regulatory role on mRNA translation. Trimethylated at Lys-318 by EEF1AKMT2. Mono-, di-, and trimethylated at Lys-36 by EEF1AKMT4; trimethylated form is predominant. Methylation by EEF1AKMT4 contributes to the fine-tuning of translation rates for a subset of tRNAs. Trimethylated at Gly-2 by EEF1AKNMT. Mono- and dimethylated at Lys-55 by EEF1AKNMT; dimethylated form is predominant.
Subcellular Location
Cell membrane, Cytoplasm, Membrane, Nucleus.
Synonyms
CCS 3 antibody
CCS3 antibody
Cervical cancer suppressor 3 antibody
chunp6927 antibody
CTCL tumor antigen antibody
EE1A1 antibody
EEF 1 antibody
EEF1A antibody
eEF1A-1 antibody
EEF1A1 antibody
ExpandImages
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Western blot analysis of eEF1A1 on different lysates with Rabbit anti-eEF1A1 antibody (ET7107-75) at 1/2,000 dilution.
Lane 1: HeLa cell lysate (10 µg/Lane)
Lane 2: MCF7 cell lysate (10 µg/Lane)
Lane 3: 293T cell lysate (10 µg/Lane)
Lane 4: Jurkat cell lysate (10 µg/Lane)
Lane 5: HepG2 cell lysate (10 µg/Lane)
Lane 6: A431 cell lysate (10 µg/Lane)
Lane 7: THP-1 cell lysate (10 µg/Lane)
Lane 8: K-562 cell lysate (10 µg/Lane)
Lane 9: C2C12 cell lysate (10 µg/Lane)
Lane 10: Neuro-2a cell lysate (10 µg/Lane)
Lane 11: NIH/3T3 cell lysate (10 µg/Lane)
Lane 12: PC-12 cell lysate (10 µg/Lane)
Lane 13: C6 cell lysate (10 µg/Lane)
Lane 14: Rat kidney tissue lysate (40 µg/Lane)
Predicted band size: 50 kDa
Observed band size: 50 kDa
Exposure time: 6 seconds; ECL: K1801;
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 (ET7107-75) 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. -
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-eEF1A1 antibody (ET7107-75) at 1/200 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 (ET7107-75) at 1/200 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. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-eEF1A1 antibody (ET7107-75) at 1/200 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 (ET7107-75) at 1/200 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. -
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-eEF1A1 antibody (ET7107-75) at 1/200 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 (ET7107-75) at 1/200 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. -
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-eEF1A1 antibody (ET7107-75) at 1/200 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 (ET7107-75) at 1/200 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. -
Immunocytochemistry analysis of HeLa cells labeling eEF1A1 with Rabbit anti-eEF1A1 antibody (ET7107-75) 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-eEF1A1 antibody (ET7107-75) 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. -
Immunocytochemistry analysis of C2C12 cells labeling eEF1A1 with Rabbit anti-eEF1A1 antibody (ET7107-75) at 1/250 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-eEF1A1 antibody (ET7107-75) at 1/250 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. -
Flow cytometric analysis of C2C12 cells labeling eEF1A1.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7107-75, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Immunocytochemistry analysis of PC-12 cells labeling eEF1A1 with Rabbit anti-eEF1A1 antibody (ET7107-75) at 1/250 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-eEF1A1 antibody (ET7107-75) at 1/250 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. -
Flow cytometric analysis of PC-12 cells labeling eEF1A1.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7107-75, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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