WB result of 6X His tag Recombinant Rabbit mAb
Primary antibody: 6X His tag Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: 4-1BB/TNFRSF9 His Tag Protein, Human 100 ng
Lane 2: Angiopoietin-1(277-498) His Tag Protein, Mouse 100 ng
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 25-33 kDa
Observed MW: 27-38 kDa
6x His tag (C-terminal) Recombinant Rabbit mAb (S-3882)
6x His tag (C-terminal) Recombinant Rabbit mAb (S-3882)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Clone Number | S-3882 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, ChIP |
| 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 | 1:1000-1:10000 | Species Independent |
| ICC | 1:500 | Species Independent |
| ChIP | 1:20-1:50 | Species Independent |
Background
A polyhistidine-tag is an amino acid motif in proteins that typically consists of at least six histidine (His) residues, often at the N- or C-terminus of the protein. The total number of histidine residues may vary in the tag from as low as two, to as high as 10 or more His residues. This antibody recognizes His-tags placed at C-terminal of fusion proteins.
Picture
Picture
Western Blot
WB result of 6X His tag Recombinant Rabbit mAb
Primary antibody: 6X His tag Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: 293T transfected with empty vector whole cell lysate 5 µg
Lane 2: 293T transfected with His-tagged fusion protein whole cell lysate 5 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 17 kDa
Observed MW: 22 kDa
Immunocytochemistry
ICC shows positive staining in MBP tag-his tag transfected 293F (top panel) and negative staining in vector-transfected 293F cells (below panel). Anti-6X His 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 293F cells were either transfected (left) or untransfected with an His-tagged human H3K27M construct (right) cross- linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication.
Parallel reactions used 6X His tag (C-terminal) Recombinant Rabbit mAb (S-3882) 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 GAPDH, RPL30, MYOD1 and
AFM in 6X His tag (C-terminal) Recombinant Rabbit mAb (S-
3882)-immunoprecipitated sample.
