Anti-GCLC Antibody Picoband™

Product Name

Anti-GCLC Antibody Picoband™

View all GCLC Antibodies

SKU/Catalog Number

PB9201

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-GCLC Antibody Picoband™ catalog # PB9201. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-GCLC Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9201)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human GCLC recombinant protein (Position: E437-N637). Human GCLC shares 94% amino acid (aa) sequence identity with both mouse and rat GCLC.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PB9201 is reactive to GCLC in Human, Mouse, Rat

Applications

PB9201 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

72 kDa

Calculated molecular weight

72.766kDa

Background of GCLC

GCLC, also named Glutamate--cysteine ligase catalytic subunit, is an enzyme that in humans is encoded by the GCLC gene. Glutamate-cysteine ligase, also known as gamma-glutamylcysteine synthetase is the first rate limiting enzyme of glutathione synthesis. The enzyme consists of two subunits, a heavy catalytic subunit and a light regulatory subunit. The gene encoding the catalytic subunit encodes a protein of 367 amino acids with a calculated molecular weight of 72.773 kDa and maps to chromosome 6p12.1. Deficiency of gamma-glutamylcysteine synthetase in human is associated with enzymopathic hemolytic anemia.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, By Heat
Flow Cytometry(Fixed), 1-3 μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of GCLC using anti-GCLC antibody (PB9201).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
Lane 2: human RT4 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human A431 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat kidney tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse kidney tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GCLC antigen affinity purified polyclonal antibody (Catalog # PB9201) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GCLC at approximately 73 kDa. The expected band size for GCLC is at 73 kDa.

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Figure 2. IHC analysis of GCLC using anti-GCLC antibody (PB9201).
GCLC was detected in a paraffin-embedded section of human colorectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-GCLC Antibody (PB9201) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. Flow Cytometry analysis of A549 cells using anti-GCLC antibody (PB9201).
Overlay histogram showing A549 cells stained with PB9201 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GCLC Antibody (PB9201, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For GCLC (Source: Uniprot.org, NCBI)

Alternative Names

EC 6.3.2.2; gamma-ECS; Gamma-glutamylcysteine synthetase; GCS heavy chain; GCS; GLCL; GLCLCGCL; glutamate--cysteine ligase catalytic subunit; glutamate-cysteine ligase, catalytic subunit GCLC GCL, GCS, GLCL, GLCLC glutamate-cysteine ligase catalytic subunit glutamate--cysteine ligase catalytic subunit|GCS heavy chain|gamma-ECS|gamma-glutamylcysteine synthetase

For more info on GCLC, check out the GCLC Infographic

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In this infographic, you will see the following information for GCLC: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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