Recombinant Anti-IFNGR1 antibody [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
Overview
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Product name
Anti-IFNGR1 antibody [EPR7866]
See all IFNGR1 primary antibodies -
Description
Rabbit monoclonal [EPR7866] to IFNGR1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 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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General notes
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 1.20 x 10 -11 M Learn more about KD -
Storage buffer
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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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7866 -
Isotype
IgG -
Research areas
Associated products
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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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab134070 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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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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Notes |
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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. |
Target
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Function
Receptor for interferon gamma. Two receptors bind one interferon gamma dimer. -
Involvement in disease
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. -
Sequence similarities
Belongs to the type II cytokine receptor family.
Contains 2 fibronectin type-III domains.
Contains 2 Ig-like C2-type (immunoglobulin-like) domains. -
Post-translational
modificationsPhosphorylated at Ser/Thr residues. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 3459 Human
- Omim: 107470 Human
- SwissProt: P15260 Human
- Unigene: 520414 Human
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Alternative names
- Antiviral Protein Type II antibody
- Antiviral protein, type 2 antibody
- AVP type II antibody
see all
Images
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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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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
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab134070 has been referenced in 7 publications.
- 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
- 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