WB result of V5 Tag Recombinant Rabbit mAb (S-609-8)
Lysate: V5 tagged fusion protein 20 ng
Primary antibody:
Lane 1: V5 Tag Recombinant Rabbit mAb at 1/5000 dilution
Lane 2: V5 Tag Recombinant Rabbit mAb at 1/10000 dilution
Lane 3: V5 Tag Recombinant Rabbit mAb at 1/50000 dilution
Lane 4: V5 Tag Recombinant Rabbit mAb at 1/100000 dilution
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
V5 Tag Recombinant Rabbit mAb (S-609-13)
V5 Tag Recombinant Rabbit mAb (S-609-13)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Immunogen | Synthetic Peptide |
| Clone Number | S-609-8 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC |
| Reactivity | Species Independent |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied. |
Dilution
| application | dilution | species |
| WB | 1:5000-1:100000 | |
| ICC | 1:500 |
Background
V5-tag is a short peptide tag for detection and purification of proteins. The V5-tag can be fused/cloned to a recombinant protein and detected in ELISA, flow cytometry, immunoprecipitation, immunofluorescence, and Western blotting with antibodies and nanobodies.
Picture
Picture
Western Blot
Immunocytochemistry
ICC shows positive staining in Histone H3-V5 tag transfected HeLa cells (top panel) and negative staining in vector-transfected HeLa cells (below panel). Anti-V5 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 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ChIP
Chromatin immunoprecipitation (ChIP) was performed on 293T cells were either untransfected (left panel) or transfected with an V5-tagged human H3 construct (right panel) cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used V5 Tag Recombinant Rabbit mAb (S-609-8), 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
V5 Tag Recombinant Rabbit mAb (S-609-8)-immunoprecipitated sample.
