Anti-Syk antibody [SYK-01]

• 产品名称

  Anti-Syk抗体[SYK-01]
  参阅全部 Syk 一抗

• 描述

  小鼠单克隆抗体[SYK-01] to Syk

• 宿主

  Mouse

• 特异性

  The antibody reacts with Protein tyrosine kinase p72Syk (Syk family tyrosine-specificphospho-transferase).

• 经测试应用

  适用于: WB, Flow Cyt, ICC/IFmore details

• 种属反应性

  与反应: Rat, Human
  

• 免疫原

  Recombinant fragment, corresponding to amino acids 5-360 of Human Syk.

• 阳性对照

• WB: Ramos cell lysate, Rat basophilic leukemia cell lysate, HeLa cell lysate; HEK-293T cell lysate, HAP1 cell lysate, THP-1 lysate; ICC/IF: THP-1 cells; Flow Cyt: Ramos cells.

• 常规说明

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

• 存储溶液

  pH: 7.40
  Preservative: 0.097% Sodium azide
  Constituent: PBS

• 纯度

  Protein A purified

• 克隆

  单克隆

• 克隆编号

  SYK-01

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab3993于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

• Entrez Gene: 6850 Human

• Entrez Gene: 25155 Rat

• Omim: 600085 Human

• SwissProt: P43405 Human

• SwissProt: Q64725 Rat

• Unigene: 371720 Human

• Unigene: 87407 Rat

• 别名

• KSYK_HUMAN antibody

• p72-Syk antibody

• p72syk antibody

• Spleen tyrosine kinase antibody

• Syk antibody

• Tyrosine protein kinase SYK antibody

• Tyrosine-protein kinase SYK antibody

• EC 2.7.10.2 antibody

• kinase Syk antibody

• KSYK antibody

• 

  Western blot - Anti-Syk antibody [SYK-01] (ab3993)

  All lanes : Anti-Syk antibody [SYK-01] (ab3993) at 1/500 dilution
  
  Lane 1 : Wild-type THP-1 lysate
  Lane 2 : Human SYK knockout THP-1 cell line (ab288700)
  
  Lysates/proteins at 20 µg per lane.
  
  Observed band size: 72 kDawhy is the actual band size different from the predicted?
  
  
  
  ab3993 was shown to react with SYK in wild-type THP-1 cells in Western blot with loss of signal observed in SYK knockout cell line ab288700. Wild-type THP-1 and SYK knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab3993 overnight at 4°C at a 1/500 dilution. Blots were incubated with secondary antibodies at 0.2µg/mL before imaging.
  
  This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Syk antibody [SYK-01] (ab3993)

  ab3993 was shown to react with SYK in wild-type THP-1 cells in immunocytochemistry with loss of signal observed in SYK knockout cell line ab288700. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab3993 at 1:1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).
  This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

• 

  Western blot - Anti-Syk antibody [SYK-01] (ab3993)

  All lanes : Anti-Syk antibody [SYK-01] (ab3993) at 1 µg/ml
  
  Lane 1 : Wild-type HEK-293T cell lysate
  Lane 2 : SYK knockout HEK-293T cell lysate
  Lane 3 : HAP1 cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Performed under reducing conditions.
  
  Observed band size: 72,73 kDawhy is the actual band size different from the predicted?
  
  
  
  

  False colour image of Western blot: Anti-Syk antibody [SYK-01] staining at 1 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab3993 was shown to bind specifically to Syk. A band was observed at 72/73 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SYK knockout cell line ab282649 (knockout cell lysate ab283048). To generate this image, wild-type and SYK knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

• 

  Western blot - Anti-Syk antibody [SYK-01] (ab3993)

  Lanes 1-2 : Anti-Syk antibody [SYK-01] (ab3993) at 2 µg/ml
  Lanes 3-4 : anti-Human Cytokeratin 18
  
  
  Lane 1 : Ramos (human Burkitt's lymphoma cell line) cell lysate
  Lane 2 : RBL (rat basophilic leukemia cell line) cell lysate
  Lanes 3-4 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
  Lane 5 : negative control
  
  Performed under non-reducing conditions.
  

• 

  Flow Cytometry - Anti-Syk antibody [SYK-01] (ab3993)

  Overlay histogram showing Ramos (Human Burkitt's lymphoma cell line) cells stained with ab3993 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab3993, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Ramos cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.