重组Anti-COX2 / Cyclooxygenase 2抗体[EPR12012] - BSA and Azide free (ab227528)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12012] to COX2 / Cyclooxygenase 2 - BSA and Azide free
- Suitable for: WB, IHC-P, IP, ICC/IF
- 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[EPR12012] - BSA and Azide free
参阅全部 COX2 / Cyclooxygenase 2 一抗 -
描述
兔单克隆抗体[EPR12012] to COX2 / Cyclooxygenase 2 - BSA and Azide free -
宿主
Rabbit -
特异性
Stimulation is required to allow detection of the COX2 protein in some cell lines and tissues. It is better to use a positive control side by side when testing.
Rat species is recommended based on IHC result, we do not guarantee WB, IP and ICC/IF for Rat. -
经测试应用
适用于: WB, IHC-P, IP, ICC/IFmore details
不适用于: Flow Cyt -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: A549, HeLa and U-87 cell lysates and mouse spleen tissue lysate. Mouse B16-F10 and Raw 264.7 whole cell lysate. Mouse retina, hippocampus, heart and kidney tissue lysate. IHC-P: Human colonic carcinoma, lung carcinoma, liver and colon tissues and rat kidney tissue lysate; mouse kidney and liver tissue. IP: A549 cell lysate.
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常规说明
ab227528 is the carrier-free version of ab179800.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR12012 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
- Alexa Fluor® 647 Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab225273)
- Alexa Fluor® 488 Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab310991)
- Alexa Fluor® 594 Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab311743)
- Alexa Fluor® 568 Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab313023)
- Alexa Fluor® 555 Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab313225)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab227528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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WB |
Use at an assay dependent concentration. Predicted molecular weight: 69 kDa.
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IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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说明 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 69 kDa. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
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 [EPR12012] (ab179800) at 1/1000 dilution
Lane 1 : RAW 264.7 Control LPS (0 ng/mL, 4 h) cell lysate
Lane 2 : RAW 264.7 Treated LPS (100 ng/mL, 4 h) cell lysate
Lane 3 : Wild-type A549 ab277305 cell lysate
Lane 4 : PTGS2 knockout A549 ab283802 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 69 kDa
Observed band size: 69 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab179800).
Western blot: Anti-PTGS2 antibody [EPR12012] (ab179800) 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, ab179800 was shown to bind specifically to PTGS2. A band was observed at 69 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in PTGS2 knockout cell line. To generate this image, wild-type and PTGS2 knockout RAW 264.7 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 [EPR12012] (ab179800) at 1/1000 dilution
Lane 1 : B16-F10 (Mouse skin melanoma) whole cell lysate
Lane 2 : Raw 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : Mouse retina tissue lysate
Lane 4 : Mouse hippocampus tissue lysate
Lane 5 : Mouse heart tissue lysate
Lane 6 : Mouse kidney tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 69 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?
Exposure time: 60 secondsBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
COX2 is expressed at a low level in Raw264.7, mouse retina, hippocampus, heart, kidney etc. (PMID: 22015457, PMID: 26001832, PMID: 23045674, PMID: 33737575).
This data was developed using the same antibody clone in a different buffer formulation (ab179800).
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All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) 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 kDaFalse colour image of Western blot: Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] 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, ab179800 was shown to bind specifically to COX2 / Cyclooxygenase 2. A band was observed at 75 kDa in wild-type A549 cell lysates with no signal observed at this size in PTGS2 knockout cell line ab280802 (knockout cell lysate ab283825). 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 fluorescent western blot (TBS-based) blocking solution 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.
This data was developed using the same antibody clone in a different buffer formulation (ab179800).
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All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/1000 dilution
Lane 1 : A549 cell lysate
Lane 2 : U-87 MG cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : PTGS2 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 69 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab179800).
Lanes 1 - 4: Merged signal (red and green). Green - ab179800 observed at 75 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab179800 was shown to react with COX2 / Cyclooxygenase 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255420 (knockout cell lysate ab263795) was used. Wild-type and COX2 / Cyclooxygenase 2 knockout samples were subjected to SDS-PAGE. ab179800 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/4000 dilution (0.125 µg/ml).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: secondary antibody only control. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800) -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/4000 dilution (0.125 µg/ml).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: secondary antibody only control. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800) -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).
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ab179800 (purified) at 1/30 immunoprecipitating COX2 in A549 whole cell lysate.
Lane 1 (input): A549 whole cell lysate (10µg)
Lane 2 (+): ab179800 + A549 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab179800 in A549 whole cell lysate.
For western blotting, HRP-conjugated anti-rabbit IgG, specific for the reduced form of IgG, was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded) analysis of human colon tissue labeling COX2 / Cyclooxygenase 2 with unpurified ab179800 at a dilution of 1/250.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded) analysis of human liver tissue labelling COX2 / Cyclooxygenase 2 with unpurified ab179800 at a dilution of 1/250.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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This IHC data was generated using the same anti-COX2 / Cyclooxygenase 2 antibody clone, EPR12012, in a different buffer formulation (cat# ab179800).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic carcinoma tissue labelling COX2 with unpurified ab179800 at a dilution of 1/250.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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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Datasheet download
Certificate of Compliance
文献 (3)
ab227528 被引用在 3 文献中.
- Qi Z et al. Effect of ginsenoside Rh2 on renal apoptosis in cisplatin-induced nephrotoxicity in vivo. Phytomedicine 61:152862 (2019). PubMed: 31048124
- Bu LJ et al. Melatonin, a novel selective ATF-6 inhibitor, induces human hepatoma cell apoptosis through COX-2 downregulation. World J Gastroenterol 23:986-998 (2017). PubMed: 28246472
- Xu K et al. Autophagy attenuates the catabolic effect during inflammatory conditions in nucleus pulposus cells, as sustained by NF-?B and JNK inhibition. Int J Mol Med 36:661-8 (2015). PubMed: 26165348