重组Anti-COX2 / Cyclooxygenase 2抗体[RM1026] (ab283574)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit recombinant multiclonal [RM1026] to COX2 / Cyclooxygenase 2
- Suitable for: ICC/IF, IP, IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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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.
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常规说明
This product is a recombinant multiclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
性能
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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 -
Concentration information loading...
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纯度
Protein A purified -
克隆
Recombinant Multiclonal -
克隆编号
RM1026 -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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Related Products
应用
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 | Ab评论 | 说明 |
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ICC/IF |
1/500.
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IP |
1/30.
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|
IHC-P |
1/500.
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|
WB |
1/1000. Predicted molecular weight: 69 kDa.
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说明 |
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ICC/IF
1/500. |
IP
1/30. |
IHC-P
1/500. |
WB
1/1000. Predicted molecular weight: 69 kDa. |
靶标
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功能
Mediates the formation of prostaglandins from arachidonate. May have a role as a major mediator of inflammation and/or a role for prostanoid signaling in activity-dependent plasticity. -
通路
Lipid metabolism; prostaglandin biosynthesis. -
序列相似性
Belongs to the prostaglandin G/H synthase family.
Contains 1 EGF-like domain. -
翻译后修饰
S-nitrosylation by NOS2 (iNOS) activates enzme activity. S-nitrosylation may take place on different Cys residues in addition to Cys-561. -
细胞定位
Microsome membrane. Endoplasmic reticulum membrane. - Information by UniProt
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数据库链接
- 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
see all -
别名
- COX 2 antibody
- COX-2 antibody
- COX2 antibody
see all
图片
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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 kDa why 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.
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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 kDa why 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.
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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 kDaBlocking 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
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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)
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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)
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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)
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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).
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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).
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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)
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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
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (3)
ab283574 被引用在 3 文献中.
- Gu G et al. Ang-(1-7)/MasR axis promotes functional recovery after spinal cord injury by regulating microglia/macrophage polarization. Cell Biosci 13:23 (2023). PubMed: 36739421
- Chen S et al. Investigating the effect of dehydromiltirone on septic AKI using a network pharmacology method, molecular docking, and experimental validation. Front Pharmacol 14:1145675 (2023). PubMed: 37007048
- Zhao W et al. The antioxidant Glycitin protects against intervertebral disc degeneration through antagonizing inflammation and oxidative stress in nucleus pulposus cells. Aging (Albany NY) 15:13693-13709 (2023). PubMed: 38019477