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Rabbit Recombinant Monoclonal GCN2 antibody. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 12 publications.


Images

Western blot - Anti-GCN2 antibody [EPR5970(2)] (AB134053), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-GCN2 antibody [EPR5970(2)] (AB134053), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-GCN2 antibody [EPR5970(2)] (AB134053), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCN2 antibody [EPR5970(2)] (AB134053), expandable thumbnail
  • Western blot - Anti-GCN2 antibody [EPR5970(2)] (AB134053), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBICC/IFFlow 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

Tested
Tested

Species
Human
Dilution info
1/100 - 1/250
Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Rat, Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000 - 1/10000
Notes

-

Not recommended
Not recommended

Species
Rat, Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/250 - 1/500
Notes

-

Not recommended
Not recommended

Species
Rat, Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100
Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000 - 1/10000

Not recommended
Not recommended

Species
Rat, Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Rat, Mouse
Dilution info
-
Notes

-

Associated Products

Select an associated product type

3 products for Alternative Product

Target data

Function

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, PubMed:32610081). 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 into 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 (PubMed:32610081). Binds uncharged tRNAs (By similarity). Required for the translational induction of protein kinase PRKCH following amino acid starvation (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).

Alternative names

GCN2, KIAA1338, EIF2AK4, eIF-2-alpha kinase GCN2, Eukaryotic translation initiation factor 2-alpha kinase 4, GCN2-like protein

Recommended products

Rabbit Recombinant Monoclonal GCN2 antibody. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 12 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR5970(2)
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle, Stable for 12 months at -20°C

Notes

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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.

13 product images

  • Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Lanes 1- 2: Merged signal (red and green). Green - 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.

    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. 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)] (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

    Lane 2: Western blot - Human EIF2AK4 (GCN2) knockout HEK-293T cell line (Human EIF2AK4 (GCN2) knockout HEK-293T cell line ab267247)

    Performed under reducing conditions.

    Predicted band size: 112 kDa, 13 kDa, 187 kDa, 55 kDa, 69 kDa

    Observed band size: 187 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling GCN2 with 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.

  • Flow Cytometry (Intracellular) - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling GCN2 with purified 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carconoma tissue sections labeling GCN2 with purified 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.

  • Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Lanes 1- 2: Merged signal (red and green). Green - 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.

    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. 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)] (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

    Lane 2: Western blot - Human EIF2AK4 (GCN2) knockout HEK-293T cell line (Human EIF2AK4 (GCN2) knockout HEK-293T cell line ab267246)

    Performed under reducing conditions.

    Predicted band size: 187 kDa

    Observed band size: 187 kDa

  • Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Lane 1: Wild-type HAP1 cell lysate (20 μg)
    Lane 2: GCN2 knockout HAP1 cell lysate (20 μg)
    Lane 3: MOLT4 cell lysate (20 μg)
    Lane 4: A549 cell lysate (20 μg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab134053 observed at 171 kDa. Red - loading control, ab18058, observed at 124 kDa.

    Unpurified ab134053 was shown to recognize GCN2 when GCN2 knockout samples were used, along with additional cross-reactive bands. Wild-type and GCN2 knockout samples were subjected to SDS-PAGE. ab134053 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.

    All lanes: Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Predicted band size: 187 kDa, 55 kDa

  • Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Blocking and diluting buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053) at 1/10000 dilution

    Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 15 µg

    Lane 2: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 187 kDa

    Observed band size: 187 kDa

  • Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Blocking and diluting buffer: 5% NFDM /TBST.

    All lanes: Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053) at 1/50000 dilution

    All lanes: MOLT-4 (Human lymphoblastic leukemia cell line) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 187 kDa

    Observed band size: 187 kDa

  • Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    All lanes: Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053) at 1/1000 dilution

    Lane 1: HeLa cell lysate at 10 µg

    Lane 2: 293T cell lysate at 10 µg

    Lane 3: MOLT4 cell lysate at 10 µg

    Lane 4: A549 cell lysate at 10 µg

    Secondary

    All lanes: HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 187 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Immunofluorescence staining of MCF-7 cells with purified 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling GCN2 with unpurified ab134053 at 1/100 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Flow Cytometry (Intracellular) - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    Overlay histogram showing HeLa cells stained with unpurified 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 (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.

  • Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053), expandable thumbnail

    Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053)

    GCN2 Western blot staining using rabbit Anti-GCN2 antibody

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/1000000 dilution.

    ab134053 is more sensitive than Anti-GCN2 antibody [EPR25345-125] ab302609 in WB testing.

    Lanes 1 - 2: Western blot - Anti-GCN2 antibody [EPR25345-125] (Anti-GCN2 antibody [EPR25345-125] ab302609) at 1/1000 dilution

    Lanes 3 - 4: Western blot - Anti-GCN2 antibody [EPR5970(2)] (ab134053) at 1/1000 dilution

    Lanes 1 and 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

    Lanes 2 and 4: 293T (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 187 kDa

    Exposure time: 60s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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