重组Anti-IFNGR1抗体[EPR7866] (ab134070)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7866] to IFNGR1
- Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-IFNGR1抗体[EPR7866]
参阅全部 IFNGR1 一抗 -
描述
兔单克隆抗体[EPR7866] to IFNGR1 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, Flow Cyt (Intra), ICC/IFmore details
不适用于: IP -
种属反应性
与反应: 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: HeLa, HEK-293T and HepG2 cell lysates. IHC-P: Human tonsil tissue. Flow Cyt (intra): HeLa cells, HEK293 cells. ICC/IF: MCF7 cells
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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 -20°C. Stable for 12 months at -20°C. -
解离常数(KD)
KD = 1.20 x 10 -11 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR7866 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab134070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 |
1/1000 - 1/10000. Detects a band of approximately 75-90 kDa (predicted molecular weight: 54 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (1) |
1/100.
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说明 |
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WB
1/1000 - 1/10000. Detects a band of approximately 75-90 kDa (predicted molecular weight: 54 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100. |
靶标
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功能
Receptor for interferon gamma. Two receptors bind one interferon gamma dimer. -
疾病相关
Defects in IFNGR1 are a cause of mendelian susceptibility to mycobacterial disease (MSMD) [MIM:209950]; also known as familial disseminated atypical mycobacterial infection. This rare condition confers predisposition to illness caused by moderately virulent mycobacterial species, such as Bacillus Calmette-Guerin (BCG) vaccine and environmental non-tuberculous mycobacteria, and by the more virulent Mycobacterium tuberculosis. Other microorganisms rarely cause severe clinical disease in individuals with susceptibility to mycobacterial infections, with the exception of Salmonella which infects less than 50% of these individuals. The pathogenic mechanism underlying MSMD is the impairment of interferon-gamma mediated immunity whose severity determines the clinical outcome. Some patients die of overwhelming mycobacterial disease with lepromatous-like lesions in early childhood, whereas others develop, later in life, disseminated but curable infections with tuberculoid granulomas. MSMD is a genetically heterogeneous disease with autosomal recessive, autosomal dominant or X-linked inheritance. -
序列相似性
Belongs to the type II cytokine receptor family.
Contains 2 fibronectin type-III domains.
Contains 2 Ig-like C2-type (immunoglobulin-like) domains. -
翻译后修饰
Phosphorylated at Ser/Thr residues. -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 3459 Human
- Omim: 107470 Human
- SwissProt: P15260 Human
- Unigene: 520414 Human
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别名
- Antiviral Protein Type II antibody
- Antiviral protein, type 2 antibody
- AVP type II antibody
see all
图片
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All lanes : Anti-IFNGR1 antibody [EPR7866] (ab134070) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : IFNGR1 knockout A549 cell lysate
Lane 3 : Wild-type HEK-293 ab259780 cell lysate
Lane 4 : IGNGR1 knockout HEK-293 ab269471 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 70-75 kDa why is the actual band size different from the predicted?Western blot: Anti-IFNGR1 antibody [EPR7866] (ab134070) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab134070 was shown to bind specifically to IFNGR1. A band was observed at 70-75 kDa in wild-type A549 cell lysates with no signal observed at this size in IFNGR1 knockout cell line. To generate this image, wild-type and IFNGR1 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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Flow cytometry overlay histogram showing wild-type HEK293 (green line) and IFNGR1 knockout HEK293 stained with ab134070 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab134070) (1x 106 in 100μl at 0.2 μg/ml (1/10000)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type HEK293 - black line, IFNGR1 knockout HEK293 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in HEK293 Fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
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All lanes : Anti-IFNGR1 antibody [EPR7866] (ab134070) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : IFNGR1 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 60-80 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab134070 observed at 60-80 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab134070 was shown to react with IFNGR1 in wild-type HEK-293 cells in western blot with loss of signal observed in IFNGR1 knockout sample. Wild-type and IFNGR1 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab134070 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 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 hour at room temperature before imaging.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IFNGR1 (red) with ab134070 at a 1/1000 dilution. Cells were fixed with 80% methanol and permeabilized with 0.1% Tween-20. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IFNGR1 antibody [EPR7866] (ab134070)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling IFNGR1 with ab134070 at 1/100 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunocytochemistry/ Immunofluorescence analysis of MCF7 (human breast adenocarcinoma epithelial cell) cells labeling IFNGR1 with purified ab134070 at 1/100 dilution (10 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL) was used as the secondary antibody only control.
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All lanes : Anti-IFNGR1 antibody [EPR7866] (ab134070) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : IFNGR1 knockout HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 54 kDa
Observed band size: 70-95 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab134070 observed at 70-95 kDa. Red - loading control ab8245 observed at 36 kDa.
ab134070 Anti-IFNGR1 antibody [EPR7866] was shown to specifically react with IFNGR1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265111 (knockout cell lysate ab257477) was used. Wild-type and IFNGR1 knockout samples were subjected to SDS-PAGE. ab134070 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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Anti-IFNGR1 antibody [EPR7866] (ab134070) at 1/1000 dilution + HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 54 kDa
Observed band size: 75-90 kDa why is the actual band size different from the predicted?
Exposure time: 60 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (8)
ab134070 被引用在 8 文献中.
- Bailey SR et al. Blockade or Deletion of IFNγ Reduces Macrophage Activation without Compromising CAR T-cell Function in Hematologic Malignancies. Blood Cancer Discov 3:136-153 (2022). PubMed: 35015685
- Wang H et al. Purine-Induced IFN-γ Promotes Uric Acid Production by Upregulating Xanthine Oxidoreductase Expression. Front Immunol 13:773001 (2022). PubMed: 35154100
- Wang B et al. New AKT-dependent mechanisms of anti-COVID-19 action of high-CBD Cannabis sativa extracts. Cell Death Discov 8:110 (2022). PubMed: 35277472
- Kuret T et al. Comprehensive transcriptome profiling of urothelial cells following TNFα stimulation in an in vitro interstitial cystitis/bladder pain syndrome model. Front Immunol 13:960667 (2022). PubMed: 36045687
- LaPlante EL et al. XDec-CHI reveals immunosuppressive interactions in pancreatic ductal adenocarcinoma. iScience 25:105249 (2022). PubMed: 36274954
- Zan J et al. Paraspeckle Promotes Hepatocellular Carcinoma Immune Escape by Sequestering IFNGR1 mRNA. Cell Mol Gastroenterol Hepatol 12:465-487 (2021). PubMed: 33667716
- Martin V et al. Met inhibition revokes IFN?-induction of PD-1 ligands in MET-amplified tumours. Br J Cancer 120:527-536 (2019). PubMed: 30723303
- Sen A et al. Rotavirus degrades multiple type interferon receptors to inhibit IFN signaling and protects against mortality from endotoxin in suckling mice. J Virol N/A:N/A (2017). PubMed: 29070687