Recombinant Anti-GCLM antibody [EPR6667] (ab126704)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)](https://www.abcam.com/ps/products/126/ab126704/Images/ab126704-10-anti-gclm-antibody-epr6667-immunohistochemistry-liver-rat.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)")
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
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-GCLM antibody [EPR6667]
See all GCLM primary antibodies -
Description
Rabbit monoclonal [EPR6667] to GCLM -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-Pmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human GCLM aa 50-150. The exact sequence is proprietary.
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Positive control
- IHC: Rat liver tissue; Mouse cardiac muscle tissue; Human bladder cancer tissue WB: HeLa, NIH/3T3, PC-12; Wild-type HAP1 whole cell lysate, HeLa cell lysate IP: HeLa cells
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General notes
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6667 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (2) |
1/1000 - 1/10000. Predicted molecular weight: 31 kDa.
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| IP |
1/10 - 1/100.
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| IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Notes |
|---|
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WB
1/1000 - 1/10000. Predicted molecular weight: 31 kDa. |
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IP
1/10 - 1/100. |
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IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Tissue specificity
In all tissues examined. Highest levels in skeletal muscle. -
Pathway
Sulfur metabolism; glutathione biosynthesis; glutathione from L-cysteine and L-glutamate: step 1/2. -
Sequence similarities
Belongs to the aldo/keto reductase family. Glutamate--cysteine ligase light chain subfamily. - Information by UniProt
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Database links
- Entrez Gene: 2730 Human
- Entrez Gene: 14630 Mouse
- Entrez Gene: 29739 Rat
- Omim: 601176 Human
- SwissProt: P48507 Human
- SwissProt: O09172 Mouse
- SwissProt: P48508 Rat
- Unigene: 315562 Human
see all -
Alternative names
- Gamma ECS regulatory subunit antibody
- Gamma-ECS regulatory subunit antibody
- Gamma-glutamylcysteine synthetase regulatory subunit antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat liver tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)](https://www.abcam.com/ps/products/126/ab126704/Images/ab126704-9-anti-gclm-antibody-epr6667-immunohistochemistry-cardiac-muscle-mouse.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cardiac muscle tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)](https://www.abcam.com/ps/products/126/ab126704/Images/ab126704-8-anti-gclm-antibody-epr6667-immunohistochemistry-bladder-cancer-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (ab126704)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder cancer tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. -
![Western blot - Anti-GCLM antibody [EPR6667] (ab126704)](https://www.abcam.com/ps/products/126/ab126704/Images/ab126704-458227-anti-gclm-antibody-epr6667-westernblot-human-mouse-rat.jpg)
Western blot - Anti-GCLM antibody [EPR6667] (ab126704)Lanes 1 & 3 : Anti-GCLM antibody [EPR6667] (ab126704) at 1/10000 dilution (Purified)
Lane 2 : Anti-GCLM antibody [EPR6667] (ab126704) at 1/10000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate
Secondary
Lanes 1 & 3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 31 kDa -
![Western blot - Anti-GCLM antibody [EPR6667] (ab126704)](https://www.abcam.com/ps/products/126/ab126704/Images/ab126704-313678-anti-gclm-antibody-epr6667-western-blot.jpg)
Western blot - Anti-GCLM antibody [EPR6667] (ab126704)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 kDaLanes 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. -
![Immunoprecipitation - Anti-GCLM antibody [EPR6667] (ab126704)](https://www.abcam.com/ps/products/126/ab126704/Images/ab126704-5-anti-gclm-antibody-epr6667-immunoprecipitation-hela-human.jpg)
Immunoprecipitation - Anti-GCLM antibody [EPR6667] (ab126704)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 -
![Anti-GCLM antibody [EPR6667] (ab126704)](https://www.abcam.com/ps/products/126/ab126704/Images/ab126704-6-benefits-of-recombinant-antibodies.png)
Anti-GCLM antibody [EPR6667] (ab126704)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (33)
ab126704 has been referenced in 33 publications.
- von Mässenhausen A et al. Dexamethasone sensitizes to ferroptosis by glucocorticoid receptor-induced dipeptidase-1 expression and glutathione depletion. Sci Adv 8:eabl8920 (2022). PubMed: 35108055
- Johnson FD et al. Characterization of a small molecule inhibitor of disulfide reductases that induces oxidative stress and lethality in lung cancer cells. Cell Rep 38:110343 (2022). PubMed: 35139387
- Tang X et al. Astragaloside-IV alleviates high glucose-induced ferroptosis in retinal pigment epithelial cells by disrupting the expression of miR-138-5p/Sirt1/Nrf2. Bioengineered 13:8240-8254 (2022). PubMed: 35302431
- Li N et al. Reciprocal regulation of NRF2 by autophagy and ubiquitin-proteasome modulates vascular endothelial injury induced by copper oxide nanoparticles. J Nanobiotechnology 20:270 (2022). PubMed: 35690781
- Liu Y et al. Cold Stress Induced Liver Injury of Mice through Activated NLRP3/Caspase-1/GSDMD Pyroptosis Signaling Pathway. Biomolecules 12:N/A (2022). PubMed: 35883482