重组Anti-TNF alpha抗体[EPR21753-109] (ab205587)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21753-109] to TNF alpha
- Suitable for: ICC/IF, WB, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat
Related conjugates and formulations
概述
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产品名称
Anti-TNF alpha抗体[EPR21753-109]
参阅全部 TNF alpha 一抗 -
描述
兔单克隆抗体[EPR21753-109] to TNF alpha -
宿主
Rabbit -
特异性
The protein level of TNF alpha in normal samples is very weak. The TNF alpha expression must be stimulated.
IL-1 beta and IL-6 could be good controls validating the increased TNF alpha level after drug treatment.
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经测试应用
适用于: ICC/IF, WB, IP, Flow Cyt (Intra)more details
不适用于: IHC-P -
种属反应性
与反应: Mouse, Rat -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Rat splenocytes treated with 20ng/ml PMA, 1µg/ml Ionomycin and 10µM BFA for 6h whole cell lysate; RAW 264.7 treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h, whole cell lysate. ICC/IF: RAW 264.7 cells. Flow: RAW 264.7 cells. IP: Rat splenocytes treated with 20ng/ml PMA, 1µg/ml Ionomycin and 10µM BFA for 6h, whole cell lysate
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常规说明
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.
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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR21753-109 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab205587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 |
---|---|---|
ICC/IF |
1/100.
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WB |
1/1000. Predicted molecular weight: 26 kDa.
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IP |
1/50.
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Flow Cyt (Intra) |
1/100.
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说明 |
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ICC/IF
1/100. |
WB
1/1000. Predicted molecular weight: 26 kDa. |
IP
1/50. |
Flow Cyt (Intra)
1/100. |
靶标
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功能
Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. -
疾病相关
Genetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis). -
序列相似性
Belongs to the tumor necrosis factor family. -
翻译后修饰
The soluble form derives from the membrane form by proteolytic processing.
The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.
O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid. -
细胞定位
Secreted and Cell membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 21926 Mouse
- Entrez Gene: 24835 Rat
- SwissProt: P06804 Mouse
- SwissProt: P16599 Rat
- Unigene: 1293 Mouse
- Unigene: 2275 Rat
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别名
- APC1 antibody
- APC1 protein antibody
- Cachectin antibody
see all
图片
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All lanes : Anti-TNF alpha antibody [EPR21753-109] (ab205587) at 1/1000 dilution (ab254360:1:1000 dilution (0.5 ug/ml)
ab259341:1:1000 dilution (0.5 ug/ml))
Lane 1 : Rat cerebral cortex tissue lysate
Lane 2 : Rat cerebellum tissue lysate
Lane 3 : Rat hippocampus tissue lysate
Lane 4 : Rat spinal cord tissue lysate
Lane 5 : Rat small intestine tissue lysate
Lane 6 : Rat kidney tissue lysate
Lane 7 : Rat skin tissue lysate
Lane 8 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 9 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml lipopolysaccharide (LPS) for 4h then add 1000ng/ml BFA for another 3h whole cell 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: 26 kDa
Observed band size: 23, 26 kDa why is the actual band size different from the predicted?
Exposure time: 180 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
Normal tissues express undetectable level of TNF alpha, IL-1 beta and IL-6 proteins.
IL-1 beta and IL-6 could be a good control validating the increased TNF alpha level after drug treatment. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (+/- treatment with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h) cells labeling TNF alpha with ab205587 at 1/100 dilution, followed by a AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 cells treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h. The Nuclear counterstain is DAPI (Blue). Tubulin was stained using an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594, ab195889) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
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All lanes : Anti-TNF alpha antibody [EPR21753-109] (ab205587) at 1/1000 dilution (ab259341 1:1000 dilution)
Lanes 1 & 3 & 5 : Untreated NR8383 (rat lung macrophage (alveolar)) whole cell lysate
Lane 2 : NR8383 (rat lung macrophage (alveolar)) treated with 100ng/ml lipopolysaccharide (LPS) for 4h then add 1000ng/ml BFA for another 3h whole cell lysate
Lane 4 : NR8383 (rat lung macrophage (alveolar)) treated with 100ng/ml lipopolysaccharide (LPS) for 3h then add 300ng/ml BFA for another 3h whole cell lysate
Lane 6 : NR8383 (rat lung macrophage (alveolar)) treated with 1000ng/ml lipopolysaccharide (LPS) for 2h whole cell 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: 26 kDa
Observed band size: 23, 26,26 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
IL-1 beta and IL-6 could be a good control validating the increased TNF alpha level after drug treatment.
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TNF alpha was immunoprecipitated from 0.35 mg rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate with ab205587 at 1/50 dilution. Western blot was performed on the immunoprecipitate using ab205587 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.
Lane 1: Rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate 10μg (input).
Lane 2: ab205587 IP in rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab205587 in rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.The MW observed is consistent with what has been described in the literature (PMID: 9933416).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h (Red) / Untreated control (Green), compared to an unlabeled control (cells without incubation with primary antibody and secondary antibody, Blue) and an isotype control- Rabbit monoclonal IgG (ab172730, Black). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077)at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-TNF alpha antibody [EPR21753-109] (ab205587) at 1/1000 dilution
Lane 1 : Untreated rat splenocytes whole cell lysate
Lane 2 : Rat splenocytes treated with 20ng/ml PMA, 1µg/ml Ionomycin and 10µM BFA for 6h whole cell lysate
Lane 3 : Untreated RAW 264.7(mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 4 : RAW 264.7 treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h 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: 26 kDaBlocking/Diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 20 seconds
The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID: 9933416)
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
Certificate of Compliance
文献 (25)
ab205587 被引用在 25 文献中.
- Aljehani AA et al. Icariin ameliorates metabolic syndrome-induced benign prostatic hyperplasia in rats. Environ Sci Pollut Res Int 29:20370-20378 (2022). PubMed: 34734339
- Zhang LQ et al. 5-HT1F Receptor Agonist Ameliorates Mechanical Allodynia in Neuropathic Pain via Induction of Mitochondrial Biogenesis and Suppression of Neuroinflammation. Front Pharmacol 13:834570 (2022). PubMed: 35308244
- Li G et al. Long non-coding RNA H19 promotes leukocyte inflammation in ischemic stroke by targeting the miR-29b/C1QTNF6 axis. CNS Neurosci Ther 28:953-963 (2022). PubMed: 35322553
- Li Y et al. Microbial and metabolic profiles of bronchopulmonary dysplasia and therapeutic effects of potential probiotics Limosilactobacillus reuteri and Bifidobacterium bifidum. J Appl Microbiol 133:908-921 (2022). PubMed: 35488863
- Binmahfouz LS et al. Piceatannol SNEDDS Attenuates Estradiol-Induced Endometrial Hyperplasia in Rats by Modulation of NF-κB and Nrf2/HO-1 Axes. Nutrients 14:N/A (2022). PubMed: 35565857