Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6667] to GCLM
- Suitable for: WB, IP, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-GCLM antibody [EPR6667]
See all GCLM primary antibodies
DescriptionRabbit monoclonal [EPR6667] to GCLM
Tested applicationsSuitable for: WB, IP, IHC-P, Flow Cytmore details
Unsuitable for: ICC
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human GCLM aa 50-150. The exact sequence is proprietary.
- HeLa, A673, PC12, A431, and K562 cell lysates; Human breast carcinoma tissue.
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.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
Our Abpromise guarantee covers the use of ab126704 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Predicted molecular weight: 31 kDa.|
|IP||1/10 - 1/100.|
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|Flow Cyt||1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Tissue specificityIn all tissues examined. Highest levels in skeletal muscle.
PathwaySulfur metabolism; glutathione biosynthesis; glutathione from L-cysteine and L-glutamate: step 1/2.
Sequence similaritiesBelongs to the aldo/keto reductase family. Glutamate--cysteine ligase light chain subfamily.
- Information by UniProt
- Gamma ECS regulatory subunit antibody
- Gamma-ECS regulatory subunit antibody
- Gamma-glutamylcysteine synthetase regulatory subunit antibody
All lanes : Anti-GCLM antibody [EPR6667] (ab126704) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : GCLM knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 31 kDa
Lanes 1 - 3: Merged signal (red and green). Green - ab126704 observed at 31 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab126704 was shown to recognize GCLM in wild-type HAP1 cells as signal was lost at the expected MW in GCLM knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GCLM knockout samples were subjected to SDS-PAGE. Ab126704 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
Overlay histogram showing HeLa cells stained with ab126704 (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 (ab126704, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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.
ab126704, at 1/50 dilution, staining GCLM in Paraffin-embedded Human breast carcinoma tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Purified ab126704 at 1/20 dilution (0.6µg) immunoprecipitating GCLM in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab126704 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126704 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 31 kDa
All lanes : Anti-GCLM antibody [EPR6667] (ab126704) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : A673 cell lysate
Lane 3 : PC12 cell lysate
Lane 4 : A431 cell lysate
Lane 5 : K562 cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Predicted band size: 31 kDa
ab126704 has been referenced in 9 publications.
- Ge C et al. Nrf2 deficiency aggravates PM2.5-induced cardiomyopathy by enhancing oxidative stress, fibrosis and inflammation via RIPK3-regulated mitochondrial disorder. Aging (Albany NY) 12:4836-4865 (2020). PubMed: 32182211
- Song CH et al. Effects of 17ß-estradiol on colorectal cancer development after azoxymethane/dextran sulfate sodium treatment of ovariectomized mice. Biochem Pharmacol 164:139-151 (2019). PubMed: 30981879
- Parsanathan R & Jain SK Hydrogen sulfide increases glutathione biosynthesis, and glucose uptake and utilisation in C2C12 mouse myotubes. Free Radic Res 52:288-303 (2018). PubMed: 29378451
- Qiu L et al. A Naturally-Occurring Dominant-Negative Inhibitor of Keap1 Competitively against Its Negative Regulation of Nrf2. Int J Mol Sci 19:N/A (2018). PubMed: 30042301
- Qiu L et al. Oncogenic Activation of Nrf2, Though as a Master Antioxidant Transcription Factor, Liberated by Specific Knockout of the Full-Length Nrf1a that Acts as a Dominant Tumor Repressor. Cancers (Basel) 10:N/A (2018). PubMed: 30562963
- Sun W et al. Proanthocyanidins Attenuation of H2O2-Induced Oxidative Damage in Tendon-Derived Stem Cells via Upregulating Nrf-2 Signaling Pathway. Biomed Res Int 2017:7529104 (2017). PubMed: 29201913
- Bakshi R et al. Neuroprotective effects of urate are mediated by augmenting astrocytic glutathione synthesis and release. Neurobiol Dis 82:574-579 (2015). PubMed: 26341543
- Zhang J et al. Comparisons of ethanol extracts of chinese propolis (poplar type) and poplar gums based on the antioxidant activities and molecular mechanism. Evid Based Complement Alternat Med 2015:307594 (2015). Mouse . PubMed: 25802536
- Jain SK et al. Hydrogen sulfide upregulates glutamate-cysteine ligase catalytic subunit, glutamate-cysteine ligase modifier subunit, and glutathione and inhibits interleukin-1ß secretion in monocytes exposed to high glucose levels. Metab Syndr Relat Disord 12:299-302 (2014). WB ; Human . PubMed: 24665821