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(Intra) |
| Clonality: | Monoclonal |
| Clone number: | JJ089-6 |
| Form: | Liquid |
| Storage condition: | 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. |
| 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(Intra) |
1:2,000-1:5,000 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 1/5,000 dilution. Lane 1: MCF7 (Human breast cancer cell) cell lysate Lane 2: MDA-MB-231 (Human breast cancer cell) cell lysate Lane 3: Jurkat (Human T-lymphoblastic cells) cell lysate Lane 4: HCT 116 (Human colon cancer cell) cell lysate Lane 5: MOLT-4 (Human acute T-Cell leukemia cell) cell lysate Lane 6: HeLa (Human cervical adenocarcinoma cell) cell lysate Lane 7: HT-29 (Human colorectal adenocarcinoma cell) cell lysate Lysates/proteins at 15 µg/Lane. Exposure time: 25 seconds; ECL: K1801 MDA-MB-231 exhibits low MGMT expression (PMID: 26420420). Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ET1701-55, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 22 kDa Observed band size: 22 kDa |
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Fig2:
Western blot analysis of MGMT on different lysates with Rabbit anti-MGMT antibody (ET1701-55) at 1/2,000 dilution. Lane 1: HAP1-parental cell lysate Lane 2: HAP1-MGMT KD cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 22 kDa Observed band size: 22 kDa Exposure time: 2 minutes 40 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 1/2,000 dilution was used in primary antibody dilution (K1803) 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:
Application: Flow Cytometry (Intra) Species: Human Sample: MCF7 (Human breast cancer cell) Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. Permeabilization: 0.1% Tween-20, 15 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. Antibody dilution buffer: 1x PBS. Primary antibody: ET1701-55(1 μg/mL, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature. Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature. |
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Fig5:
Application: Immunohistochemistry (IHC-P) Species: Human Tissue: Tonsil Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: ET1701-55, 1/200, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
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Fig6:
Application: Immunohistochemistry (IHC-P) Species: Human Tissue: Lung carcinoma Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. Primary antibody: ET1701-55, 1/50, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.