LysRS Recombinant Rabbit Monoclonal Antibody [JE40-55]
cat.: ET7109-10
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, IP, FC |
| Clonality: | Monoclonal |
| Clone number: | JE40-55 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. 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: 68 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human LysRS aa 460-597 / 597. |
| Positive control: | SiHa cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, Mouse colon tissue lysate, Mouse testis tissue lysate, Rat colon tissue lysate, Rat testis tissue lysate, MCF7, NIH/3T3, human colon tissue, human testis tissue, mouse colon tissue, mouse testis tissue, rat colon tissue, rat testis tissue. |
| Subcellular location: | Cytoplasm, cytosol, Nucleus, Cell membrane, Secreted; Mitochondrion. |
| Recommended Dilutions:
WB IHC-P IF-Cell IP FC |
1:2,000 1:200 1:100 1-2μg/sample 1:1,000 |
| Uniprot #: | SwissProt: Q15046 Human | Q99MN1 Mouse | Q5XIM7 Rat |
| Alternative names: | CMTRIB DFNB89 EC 6.1.1.6 KARS 1 KARS 2 KARS KARS1 KARS2 KIAA0070 KRS Lysine tRNA ligase Lysine--tRNA ligase LysRS Lysyl-tRNA synthetase SYK_HUMAN |
Images
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Fig1:
Western blot analysis of LysRS on different lysates with Rabbit anti-LysRS antibody (ET7109-10) at 1/2,000 dilution. Lane 1: SiHa cell lysate Lane 2: MCF7 cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: Mouse colon tissue lysate Lane 5: Mouse testis tissue lysate Lane 6: Rat colon tissue lysate Lane 7: Rat testis tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 68 kDa Observed band size: 68 kDa Exposure time: 30 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 (ET7109-10) 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:
Immunocytochemistry analysis of MCF7 cells labeling LysRS with Rabbit anti-LysRS antibody (ET7109-10) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-LysRS antibody (ET7109-10) 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 (HA601187, 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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Fig3:
Immunocytochemistry analysis of NIH/3T3 cells labeling LysRS with Rabbit anti-LysRS antibody (ET7109-10) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-LysRS antibody (ET7109-10) 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 (HA601187, 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:
Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-LysRS antibody (ET7109-10) 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 (ET7109-10) 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-LysRS antibody (ET7109-10) 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 (ET7109-10) 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. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-LysRS antibody (ET7109-10) 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 (ET7109-10) 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. |
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Fig7:
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-LysRS antibody (ET7109-10) 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 (ET7109-10) 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. |
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Fig8:
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-LysRS antibody (ET7109-10) 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 (ET7109-10) 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. |
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Fig9:
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-LysRS antibody (ET7109-10) 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 (ET7109-10) 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. |
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Fig10:
LysRS was immunoprecipitated from 0.2 mg MCF7 cell lysate with ET7109-10 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7109-10 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input) Lane 2: ET7109-10 IP in MCF7 cell lysate Lane 3: Rabbit IgG instead of ET7109-10 in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 13 seconds; ECL: K1801 |
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Fig11:
Flow cytometric analysis of MCF7 cells labeling LysRS. Cells were fixed and permeabilized. Then stained with the primary antibody (ET7109-10, 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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Fig12:
Flow cytometric analysis of NIH/3T3 cells labeling LysRS. Cells were fixed and permeabilized. Then stained with the primary antibody (ET7109-10, 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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