MGMT Recombinant Rabbit Monoclonal Antibody [JJ089-6]
cat.: ET1701-55
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IHC-P, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | JJ089-6 |
| 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: 22 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human MGMT aa 1-140 / 207. |
| Positive control: | MCF7 cell lysate, Jurkat cell lysate, HCT 116 cell lysate, HeLa cell lysate, MCF7, human tonsil tissue. |
| Subcellular location: | Nucleus. |
| Recommended Dilutions:
WB IHC-P IF-Cell FC |
1:1,000-1:5,000 1:50-1:200 1:100 1:1,000 |
| Uniprot #: | SwissProt: P16455 Human |
| Alternative names: | 6 O methylguanine DNA methyltransferase 6-O-methylguanine-DNA methyltransferase Agat AGT AI267024 EC 2.1.1.63 Methylated DNA protein cysteine methyltransferase Methylated-DNA--protein-cysteine methyltransferase Methylguanine DNA methyltransferase MGC107020 MGMT MGMT_HUMAN O 6 methylguanine DNA alkyltransferase O 6 methylguanine DNA methyltransferase O-6-methylguanine-DNA methyltransferase O-6-methylguanine-DNA-alkyltransferase |
Images
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Fig1:
Western blot analysis of MGMT on different lysates with Rabbit anti-MGMT antibody (ET1701-55) at 1/1,000 dilution. Lane 1: MCF7 cell lysate Lane 2: Jurkat cell lysate Lane 3: U-87 MG cell lysate (negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 22 kDa Observed band size: 22 kDa Exposure time: 10 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 (ET1701-55) at 1/1,000 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:
Western blot analysis of MGMT on different lysates with Rabbit anti-MGMT antibody (ET1701-55) at 1/5,000 dilution. Lane 1: MCF7 cell lysate Lane 2: HCT 116 cell lysate Lane 3: HeLa cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 22 kDa Observed band size: 22 kDa Exposure time: 25 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 (ET1701-55) at 1/5,000 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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Fig3:
Immunocytochemistry analysis of MCF7 cells labeling MGMT with Rabbit anti-MGMT antibody (ET1701-55) 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-MGMT antibody (ET1701-55) 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:
Flow cytometric analysis of MCF7 cells labeling MGMT. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-55, 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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Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-MGMT antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-55, 1/50) for 30 minutes 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.