WB result of Phospho- Zap-70 (Tyr319)/Syk (Tyr352) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho- Zap-70 (Tyr319)/Syk (Tyr352) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated Ramos whole cell lysate 20 µg
Lane 2: Serum-starved Ramos 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: 70 kDa
Observed MW: 65-75 kDa
Phospho- Zap-70 (Tyr319)/Syk (Tyr352) Recombinant Rabbit mAb (S-1252-12)
Phospho- Zap-70 (Tyr319)/Syk (Tyr352) Recombinant Rabbit mAb (S-1252-12)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho- Zap-70 (Tyr319)/Syk (Tyr352) |
| Synonyms | Tyrosine-protein kinase ZAP-70; 70 kDa zeta-chain associated protein; Syk-related tyrosine kinase; SRK; ZAP70; Tyrosine-protein kinase SYK; Spleen tyrosine kinase; p72-Syk; SYK |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Cytoskeleton, Cell membrane, Synapse |
| Accession | P43403、 P43405 |
| Clone Number | S-1252-12 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC |
| Reactivity | Hu, Rt |
| 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 | 1:1000-1:2000 | Hu, Rt |
| ICC | 1:500 | Hu |
Background
Phospho-Zap-70 (Tyr319)/Syk (Tyr352) is the dually phosphorylated, catalytically active form of the tandem SH2-bearing Syk family tyrosine kinases Zap-70 and Syk, wherein phosphorylation of Tyr319 in Zap-70 (or the homologous Tyr352 in Syk) within the interdomain B linker induces an open conformation that relieves autoinhibition, promotes recruitment to doubly phosphorylated ITAM motifs of the T-cell receptor (Zap-70) or B-cell receptor (Syk) signaling complexes, and creates docking sites for downstream adaptors such as LAT and BLNK, thereby propagating calcium flux, MAPK activation, and transcriptional programs essential for lymphocyte development, activation, and effector function.
Picture
Picture
Western Blot
WB result of Phospho- Zap-70 (Tyr319)/Syk (Tyr352) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho- Zap-70 (Tyr319)/Syk (Tyr352) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated C6 whole cell lysate 20 µg
Lane 2: C6 treated with 10 mM pervanadate for 20 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 70 kDa
Observed MW: 60-80 kDa
Dot Blot
Dot blot result of Phospho-Zap-70(Tyr319)/Syk(Tyr352) Recombinant Rabbit mAb
Lane 1: Zap-70 (Tyr319) phospho peptide 1
Lane 2: Zap-70 (Tyr319) phospho peptide 2
Lane 3: Zap-70 unmodified peptide
Primary antibody: Phospho-Zap-70(Tyr319)/Syk(Tyr352) 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 Ramos cells starve overnight then treated with pervanadate (10mM,30min) (top panel) and untreated Ramos cells (below panel). Anti- Phospho- Zap-70 (Tyr319) Syk (Tyr352) 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).
