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Product Info Summary
| SKU: | PB9201 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-GCLC Antibody Picoband®
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. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
72 kDa
Calculated molecular weight
72766 MW
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.
Application & Images
Applications
PB9201 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee
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
Positive Control
WB: human A549 whole cell, human RT4 whole cell, human HepG2 whole cell, human A431 whole cell, rat liver tissue, rat kidney tissue, mouse liver tissue, mouse kidney tissue
IHC: human colorectal cancer tissue
FCM: A549 cell
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.
Protein Target Info & Infographic
Gene/Protein Information For GCLC (Source: Uniprot.org, NCBI)
Gene Name
GCLC
Full Name
Glutamate--cysteine ligase catalytic subunit
Weight
72766 MW
Superfamily
glutamate--cysteine ligase type 3 family
Alternative Names
Glutamate--cysteine ligase catalytic subunit;6.3.2.2;GCS heavy chain;Gamma-ECS;Gamma-glutamylcysteine synthetase;GCLC;GLCL, GLCLC; GCLC GCL, GCS, GLCL, GLCLC glutamate-cysteine ligase catalytic subunit glutamate--cysteine ligase catalytic subunit|GCS heavy chain|gamma-ECS|gamma-glutamylcysteine synthetase
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on GCLC, check out the GCLC Infographic
We have 30,000+ of these available, one for each gene! Check them out.
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].
Specific Publications For Anti-GCLC Antibody Picoband® (PB9201)
Hello CJ!
PB9201 has been cited in 4 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Fucoxanthin Prevents 6-OHDA-Induced Neurotoxicity by Targeting Keap1
Zhilei Mao,Shushu Li,Lina Zhang,Mengmeng Yao,Zhu Zhou,Minjian Chen,"The mTOR/GCLc/GSH Pathway Mediates the Dose-Dependent Bidirectional Regulation of ROS Induced by TiO2 NPs in Neurogenic Cells",Oxidative Medicine and Cellular Longevity,vol.2019,Article ID 7621561,14 pages,2019.https://doi.org/10.1155/2019/7621561
Species: Human
PB9201 usage in article: APP:WB, SAMPLE: SH-SY5Y CELL, DILUTION:NA
Electroacupuncture alleviates cerebral ischemia and reperfusion injury via modulation of the ERK1/2 signaling pathway
Bone marrow mesenchymal stem cells protect against bleomycin-induced pulmonary fibrosis in rat by activating Nrf2 signaling
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Customer Q&As
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5 Customer Q&As for Anti-GCLC Antibody Picoband®
Question
We are currently using anti-GCLC antibody PB9201 for rat tissue, and we are content with the IHC results. The species of reactivity given in the datasheet says human, rat. Is it true that the antibody can work on zebrafish tissues as well?
L. Li
Verified customer
Asked: 2020-03-19
Answer
The anti-GCLC antibody (PB9201) has not been tested for cross reactivity specifically with zebrafish tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in zebrafish you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-03-19
Question
Does anti-GCLC antibody PB9201 work for IHC with liver?
R. Krishna
Verified customer
Asked: 2020-01-22
Answer
According to the expression profile of liver, GCLC is highly expressed in liver. So, it is likely that anti-GCLC antibody PB9201 will work for IHC with liver.
Boster Scientific Support
Answered: 2020-01-22
Question
Is this PB9201 anti-GCLC antibody reactive to the isotypes of GCLC?
Verified Customer
Verified customer
Asked: 2019-11-14
Answer
The immunogen of PB9201 anti-GCLC antibody is 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. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2019-11-14
Question
Does PB9201 anti-GCLC antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
B. Mitchell
Verified customer
Asked: 2018-07-03
Answer
As indicated on the product datasheet, PB9201 anti-GCLC antibody as been tested on IHC. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2018-07-03
Question
See below the WB image, lot number and protocol we used for liver using anti-GCLC antibody PB9201. Please let me know if you require anything else.
J. Thomas
Verified customer
Asked: 2014-04-18
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2014-04-18
