Rabbit Recombinant Monoclonal GCN2 antibody. Carrier free. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.
IgG
Rabbit
Constituents: PBS
Liquid
Monoclonal
IHC-P | WB | ICC/IF | Flow Cyt (Intra) | IP | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Not recommended |
Mouse | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Rat | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Rat, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Rat, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human, Rat | Dilution info - | Notes - |
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Metabolic-stress sensing protein kinase that phosphorylates the alpha subunit of eukaryotic translation initiation factor 2 (EIF2S1/eIF-2-alpha) in response to low amino acid availability (PubMed:25329545). Plays a role as an activator of the integrated stress response (ISR) required for adaptation to amino acid starvation (By similarity). EIF2S1/eIF-2-alpha phosphorylation in response to stress converts EIF2S1/eIF-2-alpha in a global protein synthesis inhibitor, leading to a global attenuation of cap-dependent translation, and thus to a reduced overall utilization of amino acids, while concomitantly initiating the preferential translation of ISR-specific mRNAs, such as the transcriptional activator ATF4, and hence allowing ATF4-mediated reprogramming of amino acid biosynthetic gene expression to alleviate nutrient depletion (By similarity). Binds uncharged tRNAs (By similarity). Involved in cell cycle arrest by promoting cyclin D1 mRNA translation repression after the unfolded protein response pathway (UPR) activation or cell cycle inhibitor CDKN1A/p21 mRNA translation activation in response to amino acid deprivation (PubMed:26102367). Plays a role in the consolidation of synaptic plasticity, learning as well as formation of long-term memory (By similarity). Plays a role in neurite outgrowth inhibition (By similarity). Plays a proapoptotic role in response to glucose deprivation (By similarity). Promotes global cellular protein synthesis repression in response to UV irradiation independently of the stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) and p38 MAPK signaling pathways (By similarity). Plays a role in the antiviral response against alphavirus infection; impairs early viral mRNA translation of the incoming genomic virus RNA, thus preventing alphavirus replication (By similarity).(Microbial infection) Plays a role in modulating the adaptive immune response to yellow fever virus infection; promotes dendritic cells to initiate autophagy and antigene presentation to both CD4(+) and CD8(+) T-cells under amino acid starvation (PubMed:24310610).
eIF-2-alpha kinase GCN2, Eukaryotic translation initiation factor 2-alpha kinase 4, GCN2-like protein, GCN2, KIAA1338, EIF2AK4
Rabbit Recombinant Monoclonal GCN2 antibody. Carrier free. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.
eIF-2-alpha kinase GCN2, Eukaryotic translation initiation factor 2-alpha kinase 4, GCN2-like protein, GCN2, KIAA1338, EIF2AK4
IgG
Rabbit
Constituents: PBS
Liquid
Monoclonal
Yes
EPR5970(2)
Affinity purification Protein A
Blue Ice
+4°C
Do Not Freeze
ab157775 is the carrier-free version of Anti-GCN2 antibody [EPR5970(2)] ab134053.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GCN2 antibody [EPR5970(2)] ab134053).
Lanes 1- 2: Merged signal (red and green). Green - Anti-GCN2 antibody [EPR5970(2)] ab134053 observed at 187 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) observed at 50 kDa.
Anti-GCN2 antibody [EPR5970(2)] ab134053 was shown to react with GCN2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human EIF2AK4 (GCN2) knockout HEK-293T cell line ab267247 (knockout cell lysate Human EIF2AK4 (GCN2) knockout HEK-293T cell lysate ab256903) was used. Wild-type HEK-293T and EIF2AK4 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Anti-GCN2 antibody [EPR5970(2)] ab134053 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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.
All lanes: Western blot - Anti-GCN2 antibody [EPR5970(2)] (Anti-GCN2 antibody [EPR5970(2)] ab134053) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: EIF2AK4 knockout HEK-293T cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 112 kDa, 13 kDa, 187 kDa, 55 kDa, 69 kDa
Observed band size: 187 kDa
Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling GCN2 with Anti-GCN2 antibody [EPR5970(2)] ab134053 at 1/250 dilution (4.0μg/ml). The cells were co-stained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 μg/ml). Cells were fixed with 100% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, a Goat anti-rabbit IgG(Alexa Fluor® 488) secondary antibody was used at 1/1000 dilution. DAPI was used as the nuclear counter stain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GCN2 antibody [EPR5970(2)] ab134053).
Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling GCN2 with purified Anti-GCN2 antibody [EPR5970(2)] ab134053 at 1/100 dilution (10 ug/ml). Cells were fixed with 4% paraformaldehyde. A Goat anti-rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Rabbit monoclonal IgG (Black) was used as the isotypre control. Cells without incubation with the primary antibody and secondary antibody (Blue) is the unlabeled control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GCN2 antibody [EPR5970(2)] ab134053).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carconoma tissue sections labeling GCN2 with purified Anti-GCN2 antibody [EPR5970(2)] ab134053 at 1/100 dilution (10 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. Tissue was counterstained with hematoxylin. PBS instead of the primary antibody was used as the negative control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GCN2 antibody [EPR5970(2)] ab134053).
This data was developed using the same antibody clone in a different buffer formation (Anti-RanBP3 antibody [EPR5088(2)] ab134052).
Lanes 1- 2: Merged signal (red and green). Green - Anti-GCN2 antibody [EPR5970(2)] ab134053 observed at 187 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) observed at 50 kDa.
Anti-GCN2 antibody [EPR5970(2)] ab134053 was shown to react with GCN2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human EIF2AK4 (GCN2) knockout HEK-293T cell line ab267246 (knockout cell lysate Human EIF2AK4 (GCN2) knockout HEK-293T cell lysate ab256902) was used. Wild-type HEK-293T and EIF2AK4 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Anti-GCN2 antibody [EPR5970(2)] ab134053 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye800®CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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.
All lanes: Western blot - Anti-GCN2 antibody [EPR5970(2)] (Anti-GCN2 antibody [EPR5970(2)] ab134053) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: EIF2AK4 knockout HEK-293T cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 187 kDa
Observed band size: 187 kDa
Immunofluorescence staining of MCF-7 cells with purified Anti-GCN2 antibody [EPR5970(2)] ab134053 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. The cells were fixed in 100% methanol. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GCN2 antibody [EPR5970(2)] ab134053).
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling GCN2 with unpurified Anti-GCN2 antibody [EPR5970(2)] ab134053 at 1/100 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GCN2 antibody [EPR5970(2)] ab134053).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Overlay histogram showing HeLa cells stained with unpurified Anti-GCN2 antibody [EPR5970(2)] ab134053 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-GCN2 antibody [EPR5970(2)] ab134053, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GCN2 antibody [EPR5970(2)] ab134053).
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