Recombinant Anti-Interferon gamma antibody [EPR23991-53] (ab267369)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23991-53] to Interferon gamma
- Suitable for: Flow Cyt (Intra), WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Interferon gamma antibody [EPR23991-53]
See all Interferon gamma primary antibodies -
Description
Rabbit monoclonal [EPR23991-53] to Interferon gamma -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WBmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 ug/mL, 6 h), BFA (5 ug/ml, last 5 h) cell lysate. Flow Cyt (intra): PBMC cells treated with cell stimulation cocktail (80nM PMA+1.34µM Ionomycin+10.6µM BFA+2uM Monensin).
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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 +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23991-53 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Dyes/Markers
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab267369 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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Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Predicted molecular weight: 19 kDa.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Predicted molecular weight: 19 kDa. |
Target
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Function
Produced by lymphocytes activated by specific antigens or mitogens. IFN-gamma, in addition to having antiviral activity, has important immunoregulatory functions. It is a potent activator of macrophages, it has antiproliferative effects on transformed cells and it can potentiate the antiviral and antitumor effects of the type I interferons. -
Tissue specificity
Released primarily from activated T lymphocytes. -
Involvement in disease
In Caucasians, genetic variation in IFNG is associated with the risk of aplastic anemia (AA) [MIM:609135]. AA is a rare disease in which the reduction of the circulating blood cells results from damage to the stem cell pool in bone marrow. In most patients, the stem cell lesion is caused by an autoimmune attack. T-lymphocytes, activated by an endogenous or exogenous, and most often unknown antigenic stimulus, secrete cytokines, including IFN-gamma, which would in turn be able to suppress hematopoiesis. -
Sequence similarities
Belongs to the type II (or gamma) interferon family. -
Post-translational
modificationsProteolytic processing produces C-terminal heterogeneity, with proteins ending alternatively at Gly-150, Met-157 or Gly-161. -
Cellular localization
Secreted. - Information by UniProt
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Database links
- Entrez Gene: 3458 Human
- Omim: 147570 Human
- SwissProt: P01579 Human
- Unigene: 856 Human
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Alternative names
- IF 1 antibody
- IFG antibody
- IFI antibody
see all
Images
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All lanes : Anti-Interferon gamma antibody [EPR23991-53] (ab267369) at 1/1000 dilution
Lane 1 : Wild-type Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 40 µg
Lane 2 : IFNG knockout Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 40 µg
Lane 3 : PTA-6967 Vehicle control + Brefeldin A (5 ug/ml, 6 h) cell lysate at 10 µg
Lane 4 : PTA-6967 Treated TPA (80 nM, 5 h), Ionomycin ab120116 (3 µM, 5 h) + Brefeldin A (5 µg/mL, 3 h) cell lysate at 10 µg
Performed under reducing conditions.
Predicted band size: 19 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Interferon gamma antibody [EPR23991-53] 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, ab267369 was shown to bind specifically to Interferon gamma. A band was observed at 28 kDa in wild-type Jurkat cell lysates with no signal observed at this size in IFNG knockout cell line ab273746 (knockout cell lysate ab275521). To generate this image, wild-type and IFNG knockout Jurkat 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-Interferon gamma antibody [EPR23991-53] (ab267369) at 1/1000 dilution
Lane 1 : Wild-type Jurkat Vehicle control: PMA (0 ng/mL, 6 h) and ionomycin (0 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 20 µg
Lane 2 : Wild-type Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 µg/mL, 6 h), BFA (5 g/ml, last 5 h) cell lysate at 20 µg
Lane 3 : IFNG knockout Jurkat Vehicle control: PMA (0 ng/mL, 6 h) and ionomycin (0 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 20 µg
Lane 4 : IFNG knockout Jurkat Treated: PMA (25 ng/mL, 6 h) and ionomycin (1 µg/mL, 6 h), BFA (5 µg/ml, last 5 h) cell lysate at 40 µg
Performed under reducing conditions.
Predicted band size: 19 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?Western blot: Anti-Interferon gamma antibody [EPR23991-53] staining at 1/1000 dilution; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab267369 was shown to bind specifically to Interferon gamma. A band was observed at 28 kDa in treated wild-type Jurkat cell lysates with no signal observed at this size in treated IFNG knockout cell line ab273746 (knockout cell lysate ab275521). To generate this image, wild-type and IFNG knockout Jurkat 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 HRP conjugated Goat anti-Rabbit (H+L) secondary antibody and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. Blot was developed with an ultra-high sensitivity ECL reagent.
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Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) treated with cell stimulation cocktail (80nM PMA+1.34uM Ionomycin+10.6uM Brefeldin A+2uM Monensin) for 6 hours cells labelling Interferon gamma with ab267369 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Cells were stained with anti-CD4 conjugated to Pacific blue. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab267369.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab267369 has been referenced in 1 publication.
- Huyan T et al. miR-221-5p and miR-186-5p Are the Critical Bladder Cancer Derived Exosomal miRNAs in Natural Killer Cell Dysfunction. Int J Mol Sci 23:N/A (2022). PubMed: 36499501