WB result of Phospho-Stat5 (Tyr694) Recombinant Rabbit mAb
Primary antibody: Phospho-Stat5 (Tyr694) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated A431 whole cell lysate 20 µg
Lane 2: A431 treated with 100 ng/ml EGF for 5 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 91 kDa
Observed MW: 90 kDa
Phospho-Stat5 (Tyr694) Recombinant Rabbit mAb (S-1183-58)
Phospho-Stat5 (Tyr694) Recombinant Rabbit mAb (S-1183-58)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho-Stat5 (Tyr694) |
| Synonyms | Signal transducer and activator of transcription 5A; STAT5A |
| Immunogen | Synthetic Peptide |
| Location | Nucleus |
| Accession | P42229 |
| Clone Number | S-1183-58 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, IP, ICFCM |
| Reactivity | Hu, Ms, Rt |
| Predicted Reactivity | Bv, Sh |
| 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 |
| Dot Blot | 1:1000 | |
| WB | 1:1000 | |
| IP | 1:50 | |
| ICC | 1:100 | |
| ICFCM | 1:200 |
Background
Stat5, which stands for Signal Transducer and Activator of Transcription 5, is a transcription factor that plays a pivotal role in cellular signaling pathways. It is a key player in cellular signaling, particularly in response to cytokine receptors such as IL-2, IL-3, IL-5, and GM-CSF. Upon binding of these cytokines to their respective receptors, a cascade of signaling events is triggered, ultimately leading to the phosphorylation of Stat5 (e.g., at Tyr694). The phosphorylated Stat5 forms dimers and translocates to the nucleus, where it acts as a transcription factor to regulate the expression of target genes.
Picture
Picture
Western Blot
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized human peripheral blood T lymphocytes, treated with CD3, CD28 beads (Right panel) or untreated (Left panel), labeling Phospho-Stat5 (Tyr694) antibody at 1/200 dilution (0.25 μg)/ (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
IP
Phospho-Stat5 (Tyr694) Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating Phospho-Stat5 (Tyr694) in 0.4 mg A431 treated with 100 ng/ml EGF for 5 minutes whole cell lysate.
Western blot was performed on the immunoprecipitate using Phospho-Stat5 (Tyr694) Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: A431 treated with 100 ng/ml EGF for 5 minutes whole cell lysate 20 µg (Input)
Lane 2: Phospho-Stat5 (Tyr694) Rabbit mAb IP in A431 treated with 100 ng/ml EGF for 5 minutes whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in A431 treated with 100 ng/ml EGF for 5 minutes whole cell lysate
Predicted MW: 91 kDa
Observed MW: 90 kDa
Dot Blot
Dot blot result of Phospho-Stat5 (Tyr694) Recombinant Rabbit mAb
Lane 1: Phospho-Stat5 (Tyr694) peptide
Lane 2: Stat5 unmodified peptide
Primary antibody: Phospho-Stat5 (Tyr694) Recombinant Rabbit mAb at 1/1000 dilution
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Immunocytochemistry
ICC analysis of A431 cells treated with EGF (100ng/mL,5min) (top panel) and A431 cells untreated with EGF (100ng/mL,5min) (below panel). Anti-Phospho-Stat5 (Tyr694) antibody was used at 1/100 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).
