Anti-Desmin antibody - Cytoskeleton Marker

• 产品名称

  Anti-COX2 / Cyclooxygenase 2抗体[RM1026]
  参阅全部 COX2 / Cyclooxygenase 2 一抗

• 描述

  兔重组multiclonal [RM1026] to COX2 / Cyclooxygenase 2

• 宿主

  Rabbit

• 经测试应用

  适用于: ICC/IF, IP, IHC-P, WBmore details
  不适用于: Flow Cyt (Intra) or IHC-Fr

• 种属反应性

  与反应: Mouse, Rat, Human
  

• 免疫原

  This product was produced with the following immunogens:
  Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
  
  Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

• 阳性对照

• WB: U-87 MG, RAW 264.7, RAW 264.7 (treated with LPS), C6, C6 (treated with LPS) cell lysates, PTGS2 (COX2 / Cyclooxygenase 2) KO A549 (Human lung carcinoma epithelial cell) cell lysate, Wild-type A549 cell lysate, U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate, MCF7 whole cell lysate. IHC-P: Human colon, Human colon carcinoma, Human liver tissues. ICC: RAW 264.7, U-87 MG cells. IP: U-87 MG, RAW 264.7 (treated with LPS) cells.

• 常规说明

  
  

  This product is a recombinant multiclonal antibody, which offers several advantages including:

  For more information see here.

  Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

• - High batch-to-batch consistency and reproducibility

• - Improved sensitivity and specificity

• - Long-term security of supply

• - Animal-free production

• 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. Avoid freeze / thaw cycle.

• 存储溶液

  pH: 7.2
  Preservative: 0.01% Sodium azide
  Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

• 浓度

• 100 µl 浓度为 0.529 mg/ml

• 40 µl 浓度为 0.529 mg/ml

• 纯度

  Protein A purified

• 克隆

  Recombinant Multiclonal

• 克隆编号

  RM1026

• 同种型

  IgG

The Abpromise guarantee

Abpromise™承诺保证使用ab283574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。

应用说明

Is unsuitable for Flow Cyt (Intra) or IHC-Fr.

• 数据库链接

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• Unigene: 292547 Mouse

• Unigene: 44369 Rat

• Entrez Gene: 5743 Human

• Entrez Gene: 19225 Mouse

• Entrez Gene: 29527 Rat

• Omim: 600262 Human

• SwissProt: P35354 Human

• SwissProt: Q05769 Mouse

• SwissProt: P35355 Rat

• Unigene: 196384 Human

• 别名

• Cyclooxygenase 2 antibody

• Cyclooxygenase 2b antibody

• Cyclooxygenase antibody

• Cyclooxygenase-2 antibody

• Cyclooxygenase2 antibody

• EC 1.14.99.1 antibody

• fj02a10 antibody

• Glucocorticoid-regulated inflammatory cyclooxygenase antibody

• Glucocorticoid-regulated inflammatory Prostaglandin G/H synthase antibody

• GRIPGHS antibody

• hCox 2 antibody

• Macrophage activation-associated marker protein P71/73 antibody

• OTTHUMP00000033524 antibody

• PES-2 antibody

• PGG/HS antibody

• PGH synthase 2 antibody

• PGH2_HUMAN antibody

• PGHS 2 antibody

• PGHS-2 antibody

• PGHS2 antibody

• PHS 2 antibody

• PHS II antibody

• PHS2 antibody

• Prostaglandin endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase) antibody

• Prostaglandin endoperoxide synthase 2 antibody

• Prostaglandin G/H synthase 2 antibody

• Prostaglandin G/H synthase 2 precursor antibody

• Prostaglandin G/H synthase and cyclooxygenase antibody

• Prostaglandin G/H synthase antibody

• Prostaglandin H2 synthase 2 antibody

• prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase) antibody

• Prostaglandin-endoperoxide synthase 2 antibody

• PTGS2 antibody

• ptgs2a antibody

• TIS10 antibody

• TIS10 protein antibody

• unp1239 antibody

• wu:fj02a10 antibody

• COX 2 antibody

• COX-2 antibody

• COX2 antibody

• 

  Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574) at 1/1000 dilution
  
  Lane 1 : Wild-type A549 cell lysate
  Lane 2 : PTGS2 knockout A549 cell lysate
  Lane 3 : U-87 MG cell lysate
  Lane 4 : MOLT-4 cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Performed under reducing conditions.
  
  Predicted band size: 69 kDa
  Observed band size: 74-76 kDawhy is the actual band size different from the predicted?
  
  
  
  

  False colour image of Western blot: Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab283574 was shown to bind specifically to COX2 / Cyclooxygenase 2. A band was observed at 74-76 kDa in wild-type A549 cell lysates with no signal observed at this size in PTGS2 knockout cell line ab280802 (knockout cell lysate ab283825). Band at 70 kDa in both wild-type and knockout samples is non-specific but exact protein is not determined. To generate this image, wild-type and PTGS2 knockout A549 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-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

• 

  Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574) at 1/10000 dilution
  
  Lane 1 : PTGS2 (COX2 / Cyclooxygenase 2) KO A549 (Human lung carcinoma epithelial cell) cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
  Lane 2 : Wild-type A549 cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
  Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
  Lane 4 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 69 kDa
  Observed band size: 74 kDawhy is the actual band size different from the predicted?
  
  
  
  

  Negative control: MCF7 (PMID: 24325753, PMID: 16997132)

  Lysates/proteins at 20 µg per lane.

  Performed under reducing conditions.

  False colour image of Western blot: Anti- COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.

  In Western blot, ab283574 was shown to bind specifically to COX2 / Cyclooxygenase 2. Target band was observed at 74 kDa in wild-type A549 cell lysates with no signal observed at this size in COX2 / Cyclooxygenase 2 knockout cell line ab280802. To generate this image, wild-type and COX2 / Cyclooxygenase 2 knockout A549 cell lysates were analyzed. 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-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

• 

  Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574) at 1/1000 dilution
  
  Lane 1 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
  Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
  Lane 3 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
  Lane 4 : RAW 264.7 treated with 1 µg/ml lipopolysaccharide (LPS) for 6h whole cell lysate
  Lane 5 : C6 (Rat glial tumor glial cell) whole cell lysate
  Lane 6 : C6 treated with 100 ng/ml lipopolysaccharide (LPS) for 4h whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
  
  Predicted band size: 69 kDa
  
  
  
  

  Blocking and diluting buffer and concentration: 5% NFDM/TBST

  Negative control: MCF7 (PMID: 24325753, PMID: 16997132)

  Lower bands could be COX-2 fragments due to proteolysis. (PMID: 32366045)

  Exposure time: Lane 1-4: 2 min Lane 5-6: 3 min

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  Immunohistochemical analysis of paraffin-embedded human colon tissue labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/500 dilution (1.058 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human colon. The section was incubated with ab283574 overnight at 4°C. Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

  Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/500 dilution (1.058 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human colon carcinoma. The section was incubated with ab283574 overnight at 4°C. Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

  Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  Immunohistochemical analysis of paraffin-embedded human liver tissue labeling COX2 / Cyclooxygenase 2 with ab283574 at 1/500 dilution (1.058 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human colon liver. The section was incubated with ab283574 overnight at 4°C. Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

  Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• 

  Immunocytochemistry/ Immunofluorescence - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/1000 dilution (0.529 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml)(Green). Confocal image showing cytoplasmic staining in RAW 264.7 cell line after treatment with lipopolysaccharide (1 µg/ml) for 6 hours is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml)(Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/ml).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell) cells labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/500 dilution (1.058 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml)(Green). Confocal image showing cytoplasmic staining in of U-87 MG cell line. Negative control: MCF7 (PMID:18199541) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml)(Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/ml).

• 

  Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  COX2 / Cyclooxygenase 2 was immunoprecipitated from 0.35 mg U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate with ab283574 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283574 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

  Lane 1: U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate 10µg

  Lane 2: ab283574 IP in U-87 MG whole cell lysate

  Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab283574 in U-87 MG whole cell lysate

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  Exposure time: 6 seconds

  Lower bands could be COX-2 fragments due to proteolysis. (PMID: 32366045)

• 

  Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)

  COX2 / Cyclooxygenase 2 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 1 µg/ml LPS for 6h whole cell lysate with ab283574 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283574 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

  Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 1ug/ml LPS for 6h whole cell lysate 10µg

  Lane 2: ab283574 IP in RAW 264.7 treated with 1 µg/ml LPS for 6h whole cell lysate

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283574 in RAW 264.7 treated with 1 µg/ml LPS for 6h whole cell lysate

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  Exposure time: 6 seconds 

• 

  Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (ab283574)