WB result of Phospho-Syk (Tyr525/526) Recombinant Rabbit mAb
Primary antibody: Phospho-Syk (Tyr525/526) Recombinant Rabbit mAb at 1/5000 dilution
Lane 1: untreated Ramos whole cell lysate 20 µg
Lane 2: Ramos starve 24 hours, then treated with 10 mM pervanadate for 30 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 72 kDa
Observed MW: 72 kDa
Phospho-Syk (Tyr525/526) Recombinant Rabbit mAb (S-1245-113)
Phospho-Syk (Tyr525/526) Recombinant Rabbit mAb (S-1245-113)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho-Syk (Tyr525/526) |
| Synonyms | Tyrosine-protein kinase SYK; Spleen tyrosine kinase; p72-Syk |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Cell membrane |
| Accession | P43405 |
| Clone Number | S-1245-113 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, IP |
| Reactivity | Hu |
| Positive Sample | Ramos starve 24 hours, then treated with 10 mM pervanadate for 30 minutes |
| Predicted Reactivity | Ms, Rt, Pg |
| 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 | 1:1000-1:5000 | Hu |
| IP | 1:50 | Hu |
| ICC | 1:500 | Hu |
Background
Phospho-Syk (Tyr525/526) refers to the phosphorylation of the tyrosine kinase Syk at the tyrosine residues 525 and 526. This post-translational modification is crucial for the activation of Syk, which plays a significant role in various signaling pathways, particularly those involved in immune cell activation and function. Syk is a non-receptor tyrosine kinase that is involved in the regulation of immune responses, including B cell receptor signaling, Fc receptor signaling, and cytokine receptor signaling. Phosphorylation at Tyr525/526 is thought to be important for the full activation of Syk and its downstream signaling events, which can include the activation of other kinases such as phospholipase C gamma 2 (PLCγ2) and the mitogen-activated protein kinases (MAPKs).
Picture
Picture
Western Blot
IP
Phospho-SYK(Tyr525/Tyr526) Recombinant Rabbit mAb (S-1245-113) at 1/50 dilution (1 µg) immunoprecipitating Phospho-SYK(Tyr525/Tyr526) in 0.4 mg Ramos+serum starvation(24) then pervanadate(10mM,30min) whole cell lysate.
Western blot was performed on the immunoprecipitate using Phospho-SYK(Tyr525/Tyr526) Recombinant Rabbit mAb (S-1245-113) at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: Ramos+serum starvation(24) then pervanadate(10mM,30min) whole cell lysate 20 µg (Input)
Lane 2: Phospho-SYK(Tyr525/Tyr526) Recombinant Rabbit mAb (S-1245-113) IP in Ramos+serum starvation(24) then pervanadate(10mM,30min) whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in Ramos+serum starvation(24) then pervanadate(10mM,30min) whole cell lysate
Predicted MW: 72 kDa
Observed MW: 72 kDa
Immunocytochemistry
ICC analysis of ramos cells serum-starved (24h) then treated with pervanadate (10mM,30min) (top panel) and ramos cells serum-starved (24h) untreated with pervanadate (10mM,30min) (below panel). Anti- Phospho-Syk (Tyr525/526) 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).
