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.
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
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.
|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.
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
This product has been referenced in:
- 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). Read more (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). Read more (PubMed: 30042301) »