Recombinant Human Interferon gamma protein (Active) is a Human Full Length protein, in the 24 to 166 aa range, expressed in HEK 293, with >=95% purity, < 0.005 EU/µg endotoxin level and suitable for SDS-PAGE, FuncS, MS, Cell Culture, HPLC.
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- APL bioengineering 7:0361082023Fluid shear stress enhances natural killer cell's cytotoxicity toward circulating tumor cells through NKG2D-mediated mechanosensing.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBing Hu,Ying Xin,Guanshuo Hu,Keming Li,Youhua TanPubMed 37575881
Key facts
- Purity
- >=95% SDS-PAGE
- Endotoxin level
- < 0.005 EU/µg
- Expression system
- HEK 293 cells
- Tags
- Tag free
- Applications
- SDS-PAGE, FuncS, MS, Cell Culture, HPLC
- Biologically active
- Yes
Amino acid sequence
Q D P Y V Q E A E N L K K Y F N A G H S D V A D N G T L F L G I L K N W K E E S D R K I M Q S Q I V S F Y F K L F K N F K D D Q S I Q K S V E T I K E D M N V K F F N S N K K K R D D F E K L T N Y S V T D L N V Q R K A I H E L I Q V M A E L S P A A K T G K R K R S Q M L F R G R R A S Q
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application SDS-PAGE | Reactivity Reacts | Dilution info - | Notes - |
Application FuncS | Reactivity Reacts | Dilution info - | Notes - |
Application MS | Reactivity Reacts | Dilution info - | Notes - |
Application Cell Culture | Reactivity Reacts | Dilution info - | Notes - |
Application HPLC | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Type II interferon produced by immune cells such as T-cells and NK cells that plays crucial roles in antimicrobial, antiviral, and antitumor responses by activating effector immune cells and enhancing antigen presentation (PubMed:16914093, PubMed:8666937). Primarily signals through the JAK-STAT pathway after interaction with its receptor IFNGR1 to affect gene regulation (PubMed:8349687). Upon IFNG binding, IFNGR1 intracellular domain opens out to allow association of downstream signaling components JAK2, JAK1 and STAT1, leading to STAT1 activation, nuclear translocation and transcription of IFNG-regulated genes. Many of the induced genes are transcription factors such as IRF1 that are able to further drive regulation of a next wave of transcription (PubMed:16914093). Plays a role in class I antigen presentation pathway by inducing a replacement of catalytic proteasome subunits with immunoproteasome subunits (PubMed:8666937). In turn, increases the quantity, quality, and repertoire of peptides for class I MHC loading (PubMed:8163024). Increases the efficiency of peptide generation also by inducing the expression of activator PA28 that associates with the proteasome and alters its proteolytic cleavage preference (PubMed:11112687). Up-regulates as well MHC II complexes on the cell surface by promoting expression of several key molecules such as cathepsins B/CTSB, H/CTSH, and L/CTSL (PubMed:7729559). Participates in the regulation of hematopoietic stem cells during development and under homeostatic conditions by affecting their development, quiescence, and differentiation (By similarity).
Alternative names
Interferon gamma, IFN-gamma, Immune interferon, IFNG
Recommended products
Recombinant Human Interferon gamma protein (Active) is a Human Full Length protein, in the 24 to 166 aa range, expressed in HEK 293, with >=95% purity, < 0.005 EU/µg endotoxin level and suitable for SDS-PAGE, FuncS, MS, Cell Culture, HPLC.
Key facts
- Purity
- >=95% SDS-PAGE
- Endotoxin level
- < 0.005 EU/µg
- Expression system
- HEK 293 cells
- Tags
- Tag free
- Applications
- SDS-PAGE, FuncS, MS, Cell Culture, HPLC
- Biologically active
- Yes
- Biological activity
- Fully biologically active determined by the dose dependent reduction in cytopathic effect of viral infection in A549 cells. ED50 for this effect is 0.30 ng/ml corresponding to a specific activity of 3.33 x 106 units/mg.
- Accession
- P01579-1
- Animal free
- Yes
- Species
- Human
- Reconstitution
- Reconstitute in PBS
- Concentration
- Storage buffer
pH: 6 - 8
Constituents: 10.26% Trehalose, 0.727% Dibasic monohydrogen potassium phosphate, 0.248% Potassium phosphate monobasic
Sequence info
Amino acid sequence
- Q D P Y V Q E A E N L K K Y F N A G H S D V A D N G T L F L G I L K N W K E E S D R K I M Q S Q I V S F Y F K L F K N F K D D Q S I Q K S V E T I K E D M N V K F F N S N K K K R D D F E K L T N Y S V T D L N V Q R K A I H E L I Q V M A E L S P A A K T G K R K R S Q M L F R G R R A S Q
- Accession
- P01579
- Protein length
- Full Length
- Predicted molecular weight
- 16.83 kDa
- Amino acids
- 24 to 166
- Nature
- Recombinant
Specifications
- Form
- Lyophilized
- Additional notes
>= 95 % HPLC.
General info
- Function
Type II interferon produced by immune cells such as T-cells and NK cells that plays crucial roles in antimicrobial, antiviral, and antitumor responses by activating effector immune cells and enhancing antigen presentation (PubMed:16914093, PubMed:8666937). Primarily signals through the JAK-STAT pathway after interaction with its receptor IFNGR1 to affect gene regulation (PubMed:8349687). Upon IFNG binding, IFNGR1 intracellular domain opens out to allow association of downstream signaling components JAK2, JAK1 and STAT1, leading to STAT1 activation, nuclear translocation and transcription of IFNG-regulated genes. Many of the induced genes are transcription factors such as IRF1 that are able to further drive regulation of a next wave of transcription (PubMed:16914093). Plays a role in class I antigen presentation pathway by inducing a replacement of catalytic proteasome subunits with immunoproteasome subunits (PubMed:8666937). In turn, increases the quantity, quality, and repertoire of peptides for class I MHC loading (PubMed:8163024). Increases the efficiency of peptide generation also by inducing the expression of activator PA28 that associates with the proteasome and alters its proteolytic cleavage preference (PubMed:11112687). Up-regulates as well MHC II complexes on the cell surface by promoting expression of several key molecules such as cathepsins B/CTSB, H/CTSH, and L/CTSL (PubMed:7729559). Participates in the regulation of hematopoietic stem cells during development and under homeostatic conditions by affecting their development, quiescence, and differentiation (By similarity).
- Sequence similarities
Belongs to the type II (or gamma) interferon family.
- Post-translational modifications
Proteolytic processing produces C-terminal heterogeneity, with proteins ending alternatively at Gly-150, Met-157 or Gly-161.
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- Ambient
- Appropriate long-term storage conditions
- Ambient
This product is an active protein and may elicit a biological response in vivo, handle with caution.
Notes
This protein is filter sterilised prior to aliquoting and lyophilisation. All aliquoting and lyophilisation steps are performed in a sterile environment
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Interferon gamma (IFN-γ) also known as type II interferon is a cytokine that plays an important role in immune response. IFN-γ has a molecular weight of about 17 kDa and is produced by T cells and natural killer (NK) cells. IFN-γ binds to the interferon gamma receptor initiating a signaling cascade that activates various genes involved in immune functions. It is expressed mainly in activated immune cells within lymphoid tissues and inflamed sites during immune responses.
- Biological function summary
This cytokine is significant in promoting macrophage activation enhancing the antigen presentation process and boosting the antimicrobial activity of phagocytes. IFN-γ is not part of a larger protein complex but works as a homodimer in signal transduction. Its production heightens the Th1 immune response by stimulating the differentiation of naïve T cells into Th1 cells which is essential for effective cellular immunity.
- Pathways
IFN-γ is integrally involved in the JAK-STAT signaling pathway alongside another critical cytokine Interleukin-12. This pathway further amplifies the immune response by regulating the expression of genes associated with cellular defense mechanisms. IFN-γ also interacts with the NF-kB pathway influencing inflammation and the activation of further immune responses. These interactions show a network of cooperativity with proteins like STAT1 and NF-kB essential for executing its biological roles.
- Associated diseases and disorders
IFN-γ is linked to autoimmune diseases such as rheumatoid arthritis and multiple sclerosis where its elevated levels can exacerbate inflammatory processes. It connects to other proteins like TNF-alpha in promoting the inflammatory cascade. Moreover lower levels of IFN-γ are associated with a heightened risk of infections like tuberculosis demonstrating its vital role in pathogen defense. Therefore understanding IFN-γ and its interactions can be key in developing therapeutic approaches against these conditions.
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Functional Studies - Recombinant Human Interferon gamma protein (Active) (ab259377)
Fully biologically active determined by the dose dependent reduction in cytopathic effect of viral infection in A549 cells. ED50 for this effect is 0.30 ng/ml corresponding to a specific activity of 3.33 x 106 units/mg.
Sandwich ELISA - Recombinant Human Interferon gamma protein (Active) (ab259377)
Wild-type A549 control cells or IP-10 knockout A549 cells (Human CXCL10 (IP10) knockout A549 cell line ab266969), grown to 40% confluency, were stimulated with Recombinant Human Interferon gamma protein (ab259377) at 100 ng/ml and Recombinant human TNF alpha protein (Recombinant human TNF alpha protein (Active) ab259410) at 10 ng/ml or vehicle control for 16 or 32 hours.
THP-1 cells, grown to 40% confluency, were stimulated with Recombinant Human Interferon gamma protein (ab259377) at 200 ng/ml and LPS at 50 ng/mL or vehicle control for 24 hours.
The concentrations of IP-10 (CXCL10) in cell culture supernatants were measured in duplicate and interpolated from the IP-10 standard curves using Human IP-10 ELISA Kit (Human IP-10 ELISA Kit ab173194). IP-10 from vehicle control samples were measured in undiluted supernatants and the treated samples were diluted 200 times. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
SDS-PAGE - Recombinant Human Interferon gamma protein (Active) (ab259377)
SDS-PAGE analysis of ab259377.
HPLC - Recombinant Human Interferon gamma protein (Active) (ab259377)
Purity: 100%
The spectrum was recorded using a 1260 Infinity II HPLC system with DAD (Agilent Technologies) and a MabPac RP column (3.0x100 mm, 4 μm, Thermo Scientific). 5 μL of purified protein was injected and the gradient run from 80 % water:TFA (99.9:0.1 v/v) and 20 % acetonitrile:water:TFA (90:9.9:0.1 v/v/v) to 20 % water:TFA (99.9:0.1 v/v) and 80 % acetonitrile:water:TFA (90:9.9:0.1 v/v/v) within 3 minutes followed by an isocratic step for another 3 min. Flow rate was 0.5 mL/min and the column compartment temperature was 50 °C.
Mass Spectrometry - Recombinant Human Interferon gamma protein (Active) (ab259377)
Mass Spectrometry analysis of ab259377 (Agilent Technologies). The mass spectrum shows the full length protein and two C-terminal truncated versions (-5 aa and -16 aa). Heterogeneity of C-terminus has been reported in literature (PAN, Yu‐Ching E., et al. "Structural characterization of human interferon γ Heterogeneity of the carboxyl terminus." European journal of biochemistry 166.1 (1987): 145-149. ).
Mass Spectrometry - Recombinant Human Interferon gamma protein (Active) (ab259377)
M + 1.4 Da (calc, mass 16833.6)
The spectrum was recorded with a 6545XT AdvanceBio LC/Q-TOF (Agilent Technologies) and a MabPac RP column (42.1x50 mm, 4 μm, Thermo Scientific). 5 μL of purified protein was injected and the gradient run from 85 % water:FA (99.9:0.1 v/v) and 15 % acetonitrile:FA (90:9.9:0.1 v/v/v) to 55 % water:FA (99.9:0.1 v/v) and 45 % acetonitrile:FA (90:9.9:0.1 v/v/v) within 3 minutes followed by an isocratic step for another 2.5 min. Flow rate was 0.4 mL/min and the column compartment temperature was 60 °C. Data was analysed and deconvoluted using the Bioconfirm software (Agilent Technologies).
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