重组Anti-Heme Oxygenase 1抗体[EP1391Y] - BSA and Azide free (ab219360)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1391Y] to Heme Oxygenase 1 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-Heme Oxygenase 1抗体[EP1391Y] - BSA and Azide free
参阅全部 Heme Oxygenase 1 一抗 -
描述
兔单克隆抗体[EP1391Y] to Heme Oxygenase 1 - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, IPmore details -
种属反应性
与反应: Mouse, Human
预测可用于: Guinea pig -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Fetal liver lysate, human liver tissue, human spleen tissue, HL-60, MCF7, A549, NIH/3T3, HEK-293, and HeLa cells. IHC-P: FFPE mouse spleen normal. Human spleen tissue. IP: Mouse spleen tissue lysate. Flow Cyt (intra): NIH/3T3 cells.
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常规说明
ab219360 is the carrier-free version of ab52947.
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.
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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
EP1391Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- PE Anti-Heme Oxygenase 1 antibody [EP1391Y] (ab305659)
- APC Anti-Heme Oxygenase 1 antibody [EP1391Y] (ab305660)
- HRP Anti-Heme Oxygenase 1 antibody [EP1391Y] (ab305661)
- Alexa Fluor® 488 Anti-Heme Oxygenase 1 antibody [EP1391Y] (ab307765)
- Alexa Fluor® 594 Anti-Heme Oxygenase 1 antibody [EP1391Y] (ab310664)
- Alexa Fluor® 555 Anti-Heme Oxygenase 1 antibody [EP1391Y] (ab312195)
- Alexa Fluor® 568 Anti-Heme Oxygenase 1 antibody [EP1391Y] (ab312683)
- Anti-Heme Oxygenase 1 antibody [EP1391Y] (ab52947)
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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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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab219360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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
靶标
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功能
Heme oxygenase cleaves the heme ring at the alpha methene bridge to form biliverdin. Biliverdin is subsequently converted to bilirubin by biliverdin reductase. Under physiological conditions, the activity of heme oxygenase is highest in the spleen, where senescent erythrocytes are sequestrated and destroyed. -
序列相似性
Belongs to the heme oxygenase family. -
细胞定位
Microsome. Endoplasmic reticulum. - Information by UniProt
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数据库链接
- Entrez Gene: 3162 Human
- Entrez Gene: 15368 Mouse
- Omim: 141250 Human
- SwissProt: P09601 Human
- SwissProt: P14901 Mouse
- Unigene: 517581 Human
- Unigene: 276389 Mouse
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别名
- 32 kD antibody
- bK286B10 antibody
- D8Wsu38e antibody
see all
图片
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All lanes : Anti-Heme Oxygenase 1 antibody [EP1391Y] (ab52947) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : HMOX1 knockout A549 cell lysate
Lane 3 : Human Spleen tissue lysate
Lane 4 : HL-60 cell lysate
Lane 5 : MCF7 cell lysate
Lane 6 : HeLa cell lysate
Lane 7 : A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 33 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab52947).
Lanes 1 - 7: Merged signal (red and green). Green - ab52947 observed at 33 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab52947 was shown to react with Heme Oxygenase 1 in wild-type A549 cells in Western blot with loss of signal observed in HMOX1 knockout cell line ab269503 (knockout cell lysate ab269665). Wild-type A549 and HMOX1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab52947 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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IHC image of ab52947 staining in mouse spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab52947, 5µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52947).
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This data was developed using ab52947, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Heme Oxygenase 1 with ab52947 at 1/2000 (0.246 ug/ml) followed by a LeicaDS9800 (Bond Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on Kupffer cells in human liver. The section was incubated with ab52947 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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This data was developed using ab52947, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Heme Oxygenase 1 with ab52947 at 1/2000 (0.246 ug/ml) followed by a LeicaDS9800 (Bond Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on mouse spleen. The section was incubated with ab52947 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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This data was developed using ab52947, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Heme Oxygenase 1 with ab52947 at 1/2000 (0.246 ug/ml) followed by a LeicaDS9800 (Bond Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on human spleen. The section was incubated with ab52947 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond Polymer Refine Detection) was used at Ready to use dilution.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Overlay histogram showing HEK293 cells stained with ab52947 (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 (ab52947, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52947).
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This data was developed using ab52947, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling Heme Oxygenase 1 with ab52947 at 1/50 dilution (1ug) (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, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Heme Oxygenase 1 was immunoprecipitated from 0.35mg mouse spleen lysate with ab52947 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab52947 at 1/1000 dilution (1 μg/mL). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/1000 dilution.
Lane 1: Mouse spleen tissue lysate 10 μg
Lane 2: Mouse spleen tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab52947 in mouse spleen lysate.Blocking and dilution 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 (ab52947).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (9)
ab219360 被引用在 9 文献中.
- Zhang J et al. Comparisons of ethanol extracts of chinese propolis (poplar type) and poplar gums based on the antioxidant activities and molecular mechanism. Evid Based Complement Alternat Med 2015:307594 (2015). Mouse . PubMed: 25802536
- Cinti A et al. Simultaneous Overexpression of Functional Human HO-1, E5NT and ENTPD1 Protects Murine Fibroblasts against TNF-a-Induced Injury In Vitro. PLoS One 10:e0141933 (2015). WB, ICC/IF ; Mouse . PubMed: 26513260
- Macak Šafranko Z et al. The effect of 17ß-estradiol on the expression of dipeptidyl peptidase III and heme oxygenase 1 in liver of CBA/H mice. J Endocrinol Invest N/A:N/A (2014). IHC-P ; Mouse . PubMed: 25432329
- Chen H et al. Nrf2 deficiency impairs the barrier function of mouse oesophageal epithelium. Gut N/A:N/A (2013). Mouse, Human . PubMed: 23676441
- Hundsdörfer C et al. UVA photoprotective properties of an artificial carotenylflavonoid hybrid molecule. Chem Res Toxicol 25:1692-8 (2012). WB . PubMed: 22799612
- Cudmore MJ et al. Loss of Akt activity increases circulating soluble endoglin release in preeclampsia: identification of inter-dependency between Akt-1 and heme oxygenase-1. Eur Heart J : (2011). PubMed: 21411816
- Zeller I et al. Lead contributes to arterial intimal hyperplasia through nuclear factor erythroid 2-related factor-mediated endothelial interleukin 8 synthesis and subsequent invasion of smooth muscle cells. Arterioscler Thromb Vasc Biol 30:1733-40 (2010). WB ; Human . PubMed: 20595649
- McCord JL et al. P2X2/3 and P2X3 receptors contribute to the metaboreceptor component of the exercise pressor reflex. J Appl Physiol 109:1416-23 (2010). PubMed: 20798273
- Wang J et al. Correlation of Nrf2, HO-1, and MRP3 in gallbladder cancer and their relationships to clinicopathologic features and survival. J Surg Res 164:e99-105 (2010). PubMed: 20828733