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9925
Phospho-Syk Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Phospho-Syk Antibody Sampler Kit #9925

Citations (1)
Confocal immunofluorescent analysis of SW620 cells (left, positive) and ACHN cells (right, negative) using Syk (D3Z1E) XP® Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Simple Western™ analysis of lysates (0.1 mg/mL) from Raji cells using Syk (D3Z1E) XP® Rabbit mAb #13198. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Simple Western™ analysis of lysates (1.0 mg/mL) from Jurkat cells using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody #2701. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
Western blot analysis of extracts from various cell lines using Syk (D3Z1E) XP® Rabbit mAb.
Western blot analysis of extracts from Jurkat cells, starved for 16 hours, and treated with 2 mM H2O2 or with calf intestinal alkaline phosphatase (CIP), using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody (upper) or control Zap-70 Antibody #2702 (lower).
Western blot analysis of extracts from Ramos cells, untreated or treated with anti-IgM, using Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb (upper) or Syk Antibody #2712 (lower).
Western blot analysis of extracts from 293T cells expressing recombinant wild-type or mutant Syk proteins, cotransfected with CD8, using Phospho-Syk (Tyr323) Antibody (upper) or Syk Antibody #2712 (lower). (Provided by Dr. Alagarsamy L. Reddi, laboratory of Dr. Hamid Band, Harvard University, Massachusetts.)
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Immunoprecipitation of Syk protein from SR cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Syk(D3Z1E) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Syk (D3Z1E) XP® Rabbit mAb
Western blot analysis of extracts from THP-1 WT (left) or SYK KO (right) using Syk (D3Z1E) XP® Rabbit mAb (upper). Membranes stained with Ponceau S for total protein normalization (lower).  These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies, as a companion to validation data generated by CST scientists.
Western blot analysis of extracts from Ramos cells, untreated or treated with anti-IgM (12 µg/ ml for 2 minutes), hydrogen peroxide (10 mM for 2 minutes) or lambda phosphatase, using Phospho-Zap-70 (Tyr319)/ Syk (Tyr352) Antibody.
Confocal immunofluorescent analysis of Ramos cells, serum-starved (overnight; left) or IgM-treated (12 ug/ml, 2 minutes; right), using Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Western blot analysis of extracts from Ramos cells, untreated or treated with anti-human-IgM (10 µg/ml), using Phospho-Syk (Tyr323) Antibody #2715 (upper) or Syk Antibody #2712 (lower).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Syk (D3Z1E) XP® Rabbit mAb.
Immunofluorescent analysis of Jurkat cells, CD3-treated (left) or untreated (right), using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody (red). Blue pseudocolor = DRAQ5 (fluorescent DNA dye).
Flow cytometric analysis of Ramos cells, untreated (blue) or treated with anti-IgM (green), using Phospho-Syk(Tyr525/526) (C87C1) Rabbit mAb, or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (PE Conjugate) #8885 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded human lymph node using Syk (D3Z1E) XP® Rabbit mAb.
Two-color flow cytometric analysis of Jurkat cells, untreated (left) or anti-CD3 activated (right), using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody and Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) mAb #9106. Anti-CD3 activation increases the intensity of label with both antibodies.
Immunohistochemical analysis of paraffin-embedded mouse spleen using Syk (D3Z1E) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Flow cytometric analysis of Jurkat cells, untreated (blue) or CD3 treated (green), using Phospho-Zap70 (Tyr319)/Syk (Tyr352) Antibody compared to a nonspecific negative control antibody (red).
Flow cytometric analysis of RL cells using Syk (D3Z1E) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.
To Purchase # 9925
Cat. # Size Qty. Price
9925T
1 Kit  (4 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-Syk (Tyr323) Antibody 2715 20 µl
  • WB
  • IP
H 72 Rabbit 
Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb 2710 20 µl
  • WB
  • IP
  • IF
  • F
H 72 Rabbit IgG
Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody 2701 20 µl
  • WB
  • IF
  • F
H 70 Zap-70, 72 Syk Rabbit 
Syk (D3Z1E) XP® Rabbit mAb 13198 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R 72 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

Phospho-Syk Sampler Kit provides an economical means to evaluate the activation status of Syk, including the phosphorylation of Tyr323, Tyr352 and Tyr525/526. The control Syk Antibody is also included. The kit contains enough primary and secondary antibodies for two Western blot experiments.

Specificity / Sensitivity

Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb detects endogenous levels of Syk protein only when phosphorylated at Tyr525/526 of human Syk or Tyr519/520 of mouse Syk. It also detects Syk protein when singly phosphorylated at Tyr526 of human Syk or Tyr520 of mouse Syk. Phospho-Syk (Tyr323) Antibody detects endogenous levels of Syk only when phosphorylated at Tyr323. Syk (D3Z1E) XP® Rabbit mAb recognizes endogenous levels of total Syk protein. All other antibodies in the kit do not cross-react with phosphorylated or nonphosphorylated forms of other related family members.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr323 of human Syk or Tyr319 of human Zap-70. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues around Asn463 of human Syk protein or residues surrounding Tyr525/526 of human Syk.

Background

Syk is a protein tyrosine kinase that plays an important role in intracellular signal transduction in hematopoietic cells (1-3). Syk interacts with immunoreceptor tyrosine-based activation motifs (ITAMs) located in the cytoplasmic domains of immune receptors (4). It couples the activated immunoreceptors to downstream signaling events that mediate diverse cellular responses, including proliferation, differentiation, and phagocytosis (4). There is also evidence of a role for Syk in nonimmune cells and investigators have indicated that Syk is a potential tumor suppressor in human breast carcinomas (5). Tyr323 is a negative regulatory phosphorylation site within the SH2-kinase linker region in Syk. Phosphorylation at Tyr323 provides a direct binding site for the TKB domain of Cbl (6,7). Tyr352 of Syk is involved in the association of PLCγ1 (8). Tyr525 and Tyr526 are located in the activation loop of the Syk kinase domain; phosphorylation at Tyr525/526 of human Syk (equivalent to Tyr519/520 of mouse Syk) is essential for Syk function (9).

Pathways

Explore pathways related to this product.

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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