M1510-3

Eftud2 Mouse Monoclonal Antibody [1-D10-A1]

cat.: M1510-3

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC, IF-Tissue
Clonality:	Monoclonal
Clone number:	1-D10-A1

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 109 kDa
Isotype:	IgG1
Immunogen:	Synthetic peptide within mouse Eftud2 aa 1-50 / 972.
Positive control:	HeLa cell lysate, Jurkat cell lysate, HEK-293 cell lysate, U-87 MG cell lysate, LNCaP cell lysate, HepG2 cell lysate, K-562 cell lysate, C6 cell lysate, PC-12 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, mouse testis tissue lysate, mouse kidney tissue lysate, rat brain tissue lysate, HeLa, human breast cancer tissue, human colon cancer tissue, human colon tissue, human ovary cancer tissue, mouse kidney tissue, mouse testis tissue, rat testis tissue, NIH/3T3.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P IF-Tissue FC	1:2,000 1:500 1:2,000 1:500 1:1,000
Uniprot #:	SwissProt: O08810 Mouse
Alternative names:	116 kDa 116 kDa U5 small nuclear ribonucleoprotein component EFTUD2 Elongation factor Tu GTP binding domain containing 2 Elongation factor Tu GTP-binding domain-containing protein 2 hSNU114 MFDGA MFDM SNRNP116 Snrp116 Snu114 SNU114 homolog U5 116KD U5 small nuclear ribonucleoprotein component U5 snRNP specific protein, 116 kD U5 snRNP specific protein, 116 kDa U5 snRNP-specific protein U5-116 kDa U5-116KD U5S1_HUMAN

Images

Fig1: Western blot analysis of Eftud2 on different lysates with Mouse anti-Eftud2 antibody (M1510-3) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: HEK-293 cell lysate
Lane 4: U-87 MG cell lysate
Lane 5: LNCaP cell lysate
Lane 6: HepG2 cell lysate
Lane 7: K-562 cell lysate
Lane 8: C6 cell lysate
Lane 9: PC-12 cell lysate
Lane 10: NIH/3T3 cell lysate
Lane 11: RAW264.7 cell lysate
Lane 12: Mouse testis tissue lysate
Lane 13: Mouse kidney tissue lysate
Lane 14: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 109 kDa
Observed band size: 120 kDa

Exposure time: 10 seconds;

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 (M1510-3) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of HeLa cells labeling Eftud2 with Mouse anti-Eftud2 antibody (M1510-3) at 1/500 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 Mouse anti-Eftud2 antibody (M1510-3) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Mouse anti-Eftud2 antibody (M1510-3) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1510-3) at 1/2,000 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 human colon cancer tissue with Mouse anti-Eftud2 antibody (M1510-3) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1510-3) at 1/2,000 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 human colon tissue with Mouse anti-Eftud2 antibody (M1510-3) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1510-3) at 1/2,000 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: Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue with Mouse anti-Eftud2 antibody (M1510-3) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1510-3) at 1/2,000 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-Eftud2 antibody (M1510-3) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1510-3) at 1/2,000 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Mouse anti-Eftud2 antibody (M1510-3) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1510-3) at 1/2,000 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.

	Fig9: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Mouse anti-Eftud2 antibody (M1510-3) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1510-3) at 1/2,000 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.
	Fig10: Immunofluorescence analysis of paraffin-embedded human breast cancer tissue labeling Eftud2 with Mouse anti-Eftud2 antibody (M1510-3) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (M1510-3, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig11: Immunofluorescence analysis of paraffin-embedded rat testis tissue labeling Eftud2 with Mouse anti-Eftud2 antibody (M1510-3) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (M1510-3, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

Fig12: Flow cytometric analysis of HeLa cells labeling Eftud2.

Cells were fixed and permeabilized. Then stained with the primary antibody (M1510-3, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Fig13: Flow cytometric analysis of NIH/3T3 cells labeling Eftud2.

Cells were fixed and permeabilized. Then stained with the primary antibody (M1510-3, 1μg/mL) (red) compared with Mouse IgG1 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-Mouse IgG Secondary antibody (HA1125) 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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