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Product Info Summary
| SKU: | A00259-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-CARS Antibody Picoband®
SKU/Catalog Number
A00259-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-CARS Antibody Picoband® catalog # A00259-1. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human. 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-CARS Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00259-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human CARS recombinant protein (Position: D510-Q748).
*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
A00259-1 is reactive to CARS in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
85 kDa
Calculated molecular weight
85473 MW
Background of CARS1
This CARS gene encodes a class 1 aminoacyl-tRNA synthetase, cysteinyl-tRNA synthetase. Each of the twenty aminoacyl-tRNA synthetases catalyzes the aminoacylation of a specific tRNA or tRNA isoaccepting family with the cognate amino acid. This gene is one of several located near the imprinted gene domain on chromosome 11p15.5, an important tumor-suppressor gene region. Alterations in this region have been associated with Beckwith-Wiedemann syndrome, Wilms tumor, rhabdomyosarcoma, adrenocortical carcinoma, and lung, ovarian and breast cancers. Alternative splicing of this gene results in multiple transcript variants.
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
A00259-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, 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
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
ELISA, 0.1-0.5μg/ml
Positive Control
WB: human Hela cell, human COLO-320 cell, human A549 cell, human PANC-1 cell, human 22RV1 cell
IHC: human placenta tissue
ICC/IF: U20S cell
FCM: A431 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of CARS using anti-CARS antibody (A00259-1).
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 50ug of sample under reducing conditions.
Lane 1: human Hela cell lysates,
Lane 2: human COLO-320 cell lysates,
Lane 3: human A549 cell lysates,
Lane 4: human PANC-1 cell lysates,
Lane 5: human 22RV1 cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-CARS antigen affinity purified polyclonal antibody (Catalog # A00259-1) 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 1:10000 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 CARS at approximately 85KD. The expected band size for CARS is at 85KD.
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Figure 2. IF analysis of CARS using anti-CARS antibody (A00259-1)
CARS was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-CARS Antibody (A00259-1) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 3. Flow Cytometry analysis of A431 cells using anti-CARS antibody (A00259-1).
Overlay histogram showing A431 cells stained with A00259-1 (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-CARS Antibody (A00259-1,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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Figure 4. IHC analysis of CARS using anti-CARS antibody (A00259-1).
CARS was detected in paraffin-embedded section of human placenta tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CARS Antibody (A00259-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Protein Target Info & Infographic
Gene/Protein Information For CARS (Source: Uniprot.org, NCBI)
Gene Name
CARS
Full Name
Cysteine--tRNA ligase, cytoplasmic
Weight
85473 MW
Superfamily
class-I aminoacyl-tRNA synthetase family
Alternative Names
Cysteine--tRNA ligase, cytoplasmic;6.1.1.16;Cysteinyl-tRNA synthetase;CysRS;CARS; CARS1 CARS, CYSRS, MCDDBH, MDBH, MGC:11246 cysteinyl-tRNA synthetase 1 cysteine--tRNA ligase, cytoplasmic|cysteine tRNA ligase 1, cytoplasmic|cysteine translase
*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 CARS, check out the CARS 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 CARS: 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-CARS Antibody Picoband® (A00259-1)
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No publications found for A00259-1
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Customer Q&As
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16 Customer Q&As for Anti-CARS Antibody Picoband®
Question
We are currently using anti-CARS antibody A00259-1 for mouse tissue, and we are well pleased with the IHC-F results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on canine tissues as well?
Verified Customer
Verified customer
Asked: 2020-04-22
Answer
The anti-CARS antibody (A00259-1) has not been tested for cross reactivity specifically with canine 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 canine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-04-22
Question
We have observed staining in rat brain. What should we do? Is anti-CARS antibody supposed to stain brain positively?
Verified Customer
Verified customer
Asked: 2020-01-16
Answer
According to literature brain does express CARS. According to Uniprot.org, CARS is expressed in endothelial cell, brain, testis, cervix, lung, cervix carcinoma, cervix carcinoma erythroleukemia, liver, among other tissues. Regarding which tissues have CARS expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 7987009
Cervix, Pubmed ID: 17974005
Cervix carcinoma, Pubmed ID: 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Liver, Pubmed ID: 24275569
Lung, Pubmed ID: 15489334
Testis, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2020-01-16
Question
Is there a BSA free version of anti-CARS antibody A00259-1 available?
Verified Customer
Verified customer
Asked: 2019-09-30
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-CARS antibody A00259-1 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2019-09-30
Question
Please see the WB image, lot number and protocol we used for liver using anti-CARS antibody A00259-1. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2019-09-05
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: 2019-09-05
Question
I was wanting to use your anti-CARS antibody for Flow Cytometry for human liver on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for human liver identification?
Verified Customer
Verified customer
Asked: 2019-05-23
Answer
As indicated on the product datasheet, A00259-1 anti-CARS antibody has been validated for ELISA, Flow Cytometry, IF, IHC-P, IHC-F, ICC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in human liver in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-05-23
Question
I appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for liver using anti-CARS antibody A00259-1. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-04-29
Answer
Thanks for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-04-29
Question
I see that the anti-CARS antibody A00259-1 works with Flow Cytometry, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-01-02
Answer
You can find protocols for Flow Cytometry on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2019-01-02
Question
I am looking for to test anti-CARS antibody A00259-1 on human liver for research purposes, then I may be interested in using anti-CARS antibody A00259-1 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2018-05-23
Answer
The products we sell, including anti-CARS antibody A00259-1, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2018-05-23
Question
I am looking for using your anti-CARS antibody for cysteinyl-trna aminoacylation studies. Has this antibody been tested with western blotting on a431 cells? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2018-02-20
Answer
Thank you for your inquiry. This A00259-1 anti-CARS antibody is tested on human hela, a431 cells. It is guaranteed to work for ELISA, Flow Cytometry, IF, IHC-P, IHC-F, ICC, WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2018-02-20
Question
I have a question about product A00259-1, anti-CARS antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2018-02-07
Answer
We do not advise storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free A00259-1 anti-CARS antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2018-02-07
Question
Will anti-CARS antibody A00259-1 work for Flow Cytometry with liver?
Verified Customer
Verified customer
Asked: 2017-10-18
Answer
According to the expression profile of liver, CARS is highly expressed in liver. So, it is likely that anti-CARS antibody A00259-1 will work for Flow Cytometry with liver.
Boster Scientific Support
Answered: 2017-10-18
Question
Is a blocking peptide available for product anti-CARS antibody (A00259-1)?
J. Bhatt
Verified customer
Asked: 2017-06-05
Answer
We do provide the blocking peptide for product anti-CARS antibody (A00259-1). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2017-06-05
Question
Is this A00259-1 anti-CARS antibody reactive to the isotypes of CARS?
L. Jones
Verified customer
Asked: 2017-03-28
Answer
The immunogen of A00259-1 anti-CARS antibody is E. coli-derived human CARS recombinant protein (Position: D510-Q748). 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: 2017-03-28
Question
Would A00259-1 anti-CARS antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
D. Johnson
Verified customer
Asked: 2016-12-30
Answer
It shows on the product datasheet, A00259-1 anti-CARS antibody as been tested on Flow Cytometry. 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: 2016-12-30
Question
My boss were well pleased with the WB result of your anti-CARS antibody. However we have been able to see positive staining in brain cytoplasm. using this antibody. Is that expected? Could you tell me where is CARS supposed to be expressed?
G. Yang
Verified customer
Asked: 2015-02-12
Answer
From literature, brain does express CARS. Generally CARS expresses in cytoplasm. Regarding which tissues have CARS expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 7987009
Cervix, Pubmed ID: 17974005
Cervix carcinoma, Pubmed ID: 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Liver, Pubmed ID: 24275569
Lung, Pubmed ID: 15489334
Testis, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2015-02-12
Question
We ordered your anti-CARS antibody for ICC on lung in a previous experiment. I am using mouse, and We want to use the antibody for WB next. I am interested in examining lung as well as testis in our next experiment. Could give a recommendation on which antibody would work the best for WB?
L. Edwards
Verified customer
Asked: 2013-02-12
Answer
I viewed the website and datasheets of our anti-CARS antibody and it seems that A00259-1 has been validated on mouse in both ICC and WB. Thus A00259-1 should work for your application. Our Boster satisfaction guarantee will cover this product for WB in mouse even if the specific tissue type has not been validated. We do have a comprehensive range of products for WB detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2013-02-12
