ET7107-75

eEF1A1 Recombinant Rabbit Monoclonal Antibody [JB44-13]

cat.: ET7107-75

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, FC, IF-Cell, IF-Tissue
Clonality:	Monoclonal
Clone number:	JB44-13

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 50 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human eEF1A1 aa 413-462 / 462.
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.
Subcellular location:	Cell membrane, Cytoplasm, Membrane, Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:2,000 1:100-1:250 1:50-1:200 1:200-1:1,000 1:1,000
Uniprot #:	SwissProt: P68104 Human \| P10126 Mouse \| P62630 Rat
Alternative names:	CCS 3 CCS3 Cervical cancer suppressor 3 chunp6927 CTCL tumor antigen EE1A1 EEF 1 EEF1A eEF1A-1 EEF1A1 EF-1-alpha-1 EF-Tu EF1A EF1a like protein EF1A1_HUMAN Elongation factor 1 alpha subunit Elongation factor 1-alpha 1 Elongation factor Tu Eukaryotic elongation factor 1 A-1 Eukaryotic translation elongation factor 1 alpha 1 Eukaryotic translation elongation factor 1 alpha 1 like 14 Glucocorticoid receptor AF 1 specific elongation factor GRAF 1EF HNGC:16303 ik:tdsubc_2a3 ik:tdsubc_2b3 LENG7 Leukocyte receptor cluster (LRC) member 7 Leukocyte receptor cluster member 7 Prostate tumor inducing protein 1 PTI1 tdsubc_2a3 Translation elongation factor 1 alpha 1 like 14 wu:fa91c07 wu:fa94b03 wu:fi13b09 xx:tdsubc_2a3 xx:tdsubc_2b3

Images

Fig1: 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.

Fig2: 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.

	Fig3: 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.
	Fig4: 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.
	Fig5: 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.

	Fig6: 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.
	Fig7: 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.
	Fig8: 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).

Fig9: 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.

Fig10: 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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