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: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-COX2 / Cyclooxygenase 2 antibody [RM1026]
COX2 / Cyclooxygenase 2 一次抗体 製品一覧 -
製品の詳細
Rabbit recombinant multiclonal [RM1026] to COX2 / Cyclooxygenase 2 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WBmore details
適用なし: IHC-Fr -
種交差性
交差種: 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. -
ポジティブ・コントロール
- 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. FC(Intra): RAW 264.7, U-87 MG cells. IP: U-87 MG, RAW 264.7 (treated with LPS) cells.
-
特記事項
This product is a recombinant monoclonal 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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
Recombinant Multiclonal -
クローン名
RM1026 -
アイソタイプ
IgG -
研究分野
関連製品
-
Compatible Secondaries
-
Isotype control
-
Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab283574の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
Flow Cyt (Intra) |
1/50.
|
|
ICC/IF |
1/500.
|
|
IP |
1/30.
|
|
IHC-P |
1/500.
|
|
WB |
1/1000. Predicted molecular weight: 69 kDa.
|
特記事項 |
---|
Flow Cyt (Intra)
1/50. |
ICC/IF
1/500. |
IP
1/30. |
IHC-P
1/500. |
WB
1/1000. Predicted molecular weight: 69 kDa. |
ターゲット情報
-
機能
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. -
パスウェイ
Lipid metabolism; prostaglandin biosynthesis. -
配列類似性
Belongs to the prostaglandin G/H synthase family.
Contains 1 EGF-like domain. -
翻訳後修飾
S-nitrosylation by NOS2 (iNOS) activates enzme activity. S-nitrosylation may take place on different Cys residues in addition to Cys-561. -
細胞内局在
Microsome membrane. Endoplasmic reticulum membrane. - Information by UniProt
-
参照データベース
- 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 -
別名
- COX 2 antibody
- COX-2 antibody
- COX2 antibody
see all
画像
-
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.
-
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.
-
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
-
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)
-
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)
-
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)
-
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).
-
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).
-
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 1 µg/ml LPS for 6h (Red) / Untreated control (Green) cells labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/500 dilution (0.1µg) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/500 dilution was used as the secondary antibody.
-
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell)(Left) / U-87 MG (human glioblastoma-astrocytoma epithelial cell)(Right) cells labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/500 dilution was used as the secondary antibody.
-
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)
-
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
プロトコール
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.
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (2)
ab283574 は 2 報の論文で使用されています。
- 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