Tested applications:WBIHC-PIF/ICCIPChIPChIP-seqRIPFCFC(Intra)ELISAMeDIPNucleotide ArrayDBFACSCoIPCUT&TagmeRIPInhibitionReactivity:Human, Mouse, Rat
Western blot analysis of lysates from C6 cells using Phospho-SMAD1-S463/S465+SMAD5-S463/S465+SMAD9-S465/S467 Rabbit mAb (AP1518) at 1:1000 dilution. C6 cells were treated by TGF beta 3(10ng/ml) and MG132(2 μM) at 37℃ for 20 minutes.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.
Immunohistochemistry analysis of paraffin-embedded Human lung cancer tissue using Phospho-SMAD1-S463/S465+SMAD5-S463/S465+SMAD9-S465/S467 Rabbit mAb (AP1518) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.
Immunohistochemistry analysis of paraffin-embedded Rat liver tissue using Phospho-SMAD1-S463/S465+SMAD5-S463/S465+SMAD9-S465/S467 Rabbit mAb (AP1518) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.
Confocal imaging of HeLa cells (treated with BMP2) and HeLa cells (untreated) using Phospho-SMAD1-S463/S465+SMAD5-S463/S465+SMAD9-S465/S467 Rabbit mAb (AP1518, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
| Product name | Phospho-SMAD1-S463/S465+SMAD5-S463/S465+SMAD9-S465/S467 Rabbit mAb |
|---|---|
| Catalog No. | AP1518 |
| Host species | Rabbit |
| Purification method | Affinity purification |
| Isotype | IgG |
| CloneNo. | ARC59007 |
| Immunogen | A synthetic phosphorylated peptide around S463/S465//S467 of human SMAD1/SMAD5/SMAD9. |
|---|---|
| Sequence | PISSVS |
| Gene ID | |
| Swiss Prot | |
| Synonyms | BSP1; MADH1; MADR1; JV4-1; DWFC; JV5-1; MADH5; MADH6; MADH9; SMAD8; Phospho-SMAD1-S463/S465+SMAD5-S463/S465+SMAD9-S465/S467 |
| Calculated MW | 52kDa |
| Observed MW | 60kDa |
| Reactivity | Human, Mouse, Rat |
|---|---|
| Tested applications | WBIHC-PIF/ICCIPChIPChIP-seqRIPFCFC(Intra)ELISAMeDIPNucleotide ArrayDBFACSCoIPCUT&TagmeRIPInhibition |
| Recommended dilution |
|
| Storage buffer | Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3. |
| Key application | Western blotting Immunohistochemistry Immunofluorescence |
| Positive samples | C6 treated by TGF beta 3 and MG132 |
| Cellular location | Cytoplasm, Nucleus. |
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