PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300
12 months from date of receipt / reconstitution, -20 °C as supplied
| 应用 | 稀释度 |
|---|---|
| WB | 1:5000-1:100000 |
| IP | 1:100 |
| ICC | 1:500 |
| ICFCM | 1:5000 |
| ChIP | 1:20-1:50 |
A myc tag is a polypeptide protein tag derived from the c-myc gene product that can be added to a protein using recombinant DNA technology. It can be used for affinity chromatography, then used to separate recombinant, overexpressed protein from wild type protein expressed by the host organism. It can also be used in the isolation of protein complexes with multiple subunits.
WB result of Myc tag Rabbit mAb
Primary antibody: Myc tag Rabbit mAb at 1/1000 dilution
Lane 1: 293F whole cell lysate 5 µg
Lane 2: 293F transfected with Myc-GFP fusion protein whole cell lysate 20µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 27 kDa
Observed MW: 27 kDa
Exposure time: 30 s
WB result of Myc tag Rabbit mAb
Primary antibody: Myc tag Rabbit mAb at 1/1000 dilution
Lane 1: 293T whole cell lysate 5 µg
Lane 2: 293T transfected with Myc-Histone H3.3 fusion protein whole cell lysate 20µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 13 kDa
Observed MW: 18 kDa
Exposure time: 60 s
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized H3.3 (G34W mutant)-Myc-His transfected HeLa (Human cervix adenocarcinoma epithelial cell) labelling Myc tag antibody at 1/5000 (0.01 μg) dilution/ (right panel) compared with a Rabbit IgG Isotype Control / (left panel). Goat Anti-Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody. Then cells were stained with His Tag - Alexa Fluor® 647 separately.
Myc tag Rabbit mAb at 1/100 dilution (0.5 µg) immunoprecipitating Myc tag in 0.4 mg 293F transfected with Myc-GFP fusion protein whole cell lysate.
Western blot was performed on the immunoprecipitate using Myc tag Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1: 293F transfected with Myc-GFP fusion protein whole cell lysate 5 µg (Input)
Lane 2: Myc tag Rabbit mAb IP in 293F transfected with Myc-GFP fusion protein whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in 293F transfected with Myc-GFP fusion protein whole cell lysate
Predicted MW: 27 kDa
Observed MW: 27 kDa
ICC shows positive staining in Myc-Histone H3.3 transfected HeLa cells. Anti-Myc tag antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).
Negative control: ICC shows negative staining in vector-transfected HeLa cells. Anti-Myc tag antibody was used at 1/500 dilution and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).
Chromatin immunoprecipitation (ChIP) was performed on 293T cells were either untransfected (left panel) or transfected with an Myc - tagged human H3 construct (right panel) cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used Myc tag Recombinant Rabbit mAb (S-114-13), Histone H3 Recombinant Rabbit mAb (SDT-266-44) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation.
Post - immunoprecipitation, both samples were washed, eluted, and cross - links reversed. Purified DNA was analyzed by qPCR.
qPCR (%input: immunoprecipitated DNA/input DNA)
showed the enrichment of RPL30, MYOD1 and SAT-2 in
Myc tag Recombinant Rabbit mAb
(S-114-13)-immunoprecipitated sample.
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