Product Info Summary
SKU: | PB9987 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-ARF6 Antibody Picoband®
SKU/Catalog Number
PB9987
PB1040 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ARF6 Antibody Picoband® catalog # PB9987. Tested in Flow Cytometry, IF, IHC, ICC, 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-ARF6 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9987)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human ARF6 recombinant protein (Position: G2-S175). Human ARF6 shares 100% amino acid (aa) sequence identity with both mouse and rat ARF6.
*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
PB9987 is reactive to ARF6 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
20 kDa
Calculated molecular weight
20082 MW
Background of ARF6
ADP-ribosylation factor 6 (ARF6) is a member of the human ARF gene family, which is part of the RAS superfamily. The ARF genes encode small guanine nucleotide-binding proteins that stimulate the ADP-ribosyltransferase activity of cholera toxin and play a role in vesicular trafficking and as activators of phospholipase D. The product of this gene is localized to the plasma membrane, and regulates vesicular trafficking, remodelling of membrane lipids, and signaling pathways that lead to actin remodeling. A pseudogene of this gene is located on chromosome 7.
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
PB9987 is guaranteed for 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, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: rat kidney tissue, mouse kidney tissue, MCF-7 whole cell
IHC: human melanoma tissue, rat cardiac muscle tissue
ICC/IF: A431 cell
FCM: SiHa cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of ARF6 using anti-ARF6 antibody (PB9987).
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: rat kidney tissue lysates,
Lane 2: mouse kidney tissue lysates,
Lane 3: MCF-7 whole 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-ARF6 antigen affinity purified polyclonal antibody (Catalog # PB9987) 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 ARF6 at approximately 20KD. The expected band size for ARF6 is at 20D.
Click image to see more details
Figure 2. Flow Cytometry analysis of SiHa cells using anti-ARF6 antibody (PB9987).
Overlay histogram showing SiHa cells stained with PB9987 (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-ARF6 Antibody (PB9987,1μg/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.
Click image to see more details
Figure 3. IHC analysis of ARF6 using anti-ARF6 antibody (PB9987).
ARF6 was detected in paraffin-embedded section of human melanoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ARF6 Antibody (PB9987) 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.
Click image to see more details
Figure 4. IHC analysis of ARF6 using anti-ARF6 antibody (PB9987).
ARF6 was detected in paraffin-embedded section of rat cardiac muscle tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ARF6 Antibody (PB9987) 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.
Click image to see more details
Figure 5. IF analysis of ARF6 using anti-ARF6 antibody (PB9987).
ARF6 was detected in immunocytochemical section of A431 cells. 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-ARF6 Antibody (PB9987) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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.
Protein Target Info & Infographic
Gene/Protein Information For ARF6 (Source: Uniprot.org, NCBI)
Gene Name
ARF6
Full Name
ADP-ribosylation factor 6
Weight
20082 MW
Superfamily
small GTPase superfamily
Alternative Names
ADP-ribosylation factor 6;ARF6; ARF6 ADP ribosylation factor 6 ADP-ribosylation factor 6
*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 ARF6, check out the ARF6 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 ARF6: 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-ARF6 Antibody Picoband® (PB9987)
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Customer Q&As
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17 Customer Q&As for Anti-ARF6 Antibody Picoband®
Question
My team were satisfied with the WB result of your anti-ARF6 antibody. However we have seen positive staining in esophagus squamous epithelium cytoplasm using this antibody. Is that expected? Could you tell me where is ARF6 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2020-04-09
Answer
From what I have seen in literature, esophagus squamous epithelium does express ARF6. Generally ARF6 expresses in cytoplasm, cytosol. Regarding which tissues have ARF6 expression, here are a few articles citing expression in various tissues:
Leukocyte, Pubmed ID: 11062263
Liver, Pubmed ID: 24275569
Skin, and Uterus, Pubmed ID: 15489334
Trachea, Pubmed ID: 14702039
Umbilical cord blood, Pubmed ID: 9653160
Boster Scientific Support
Answered: 2020-04-09
Question
We are currently using anti-ARF6 antibody PB9987 for human tissue, and we are well pleased with the Flow Cytometry results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on zebrafish tissues as well?
Verified Customer
Verified customer
Asked: 2020-02-07
Answer
The anti-ARF6 antibody (PB9987) has not been validated 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: 2020-02-07
Question
I would like to test anti-ARF6 antibody PB9987 on rat liver for research purposes, then I may be interested in using anti-ARF6 antibody PB9987 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Z. Mitchell
Verified customer
Asked: 2020-01-28
Answer
The products we sell, including anti-ARF6 antibody PB9987, 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: 2020-01-28
Question
Do you have a BSA free version of anti-ARF6 antibody PB9987 available?
Verified Customer
Verified customer
Asked: 2020-01-13
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-ARF6 antibody PB9987 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: 2020-01-13
Question
I was wanting to use your anti-ARF6 antibody for WB for rat liver on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for rat liver identification?
M. Krishna
Verified customer
Asked: 2019-11-19
Answer
As indicated on the product datasheet, PB9987 anti-ARF6 antibody has been validated for Flow Cytometry, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in rat liver in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-11-19
Question
Thank you for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for liver using anti-ARF6 antibody PB9987. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-10-29
Answer
Thank you 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: 2019-10-29
Question
We have tried in the past anti-ARF6 antibody for WB on skin uterus a few months ago. I am using rat, and We want to use the antibody for Flow Cytometry next. We want examining skin uterus as well as liver in our next experiment. Could you please give me some suggestion on which antibody would work the best for Flow Cytometry?
Verified Customer
Verified customer
Asked: 2019-10-07
Answer
I have checked the website and datasheets of our anti-ARF6 antibody and I see that PB9987 has been tested on rat in both WB and Flow Cytometry. Thus PB9987 should work for your application. Our Boster satisfaction guarantee will cover this product for Flow Cytometry in rat even if the specific tissue type has not been validated. We do have a comprehensive range of products for Flow Cytometry detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2019-10-07
Question
Does anti-ARF6 antibody PB9987 work on canine Flow Cytometry with leukocyte?
Verified Customer
Verified customer
Asked: 2019-06-05
Answer
Our lab technicians have not validated anti-ARF6 antibody PB9987 on canine. You can run a BLAST between canine and the immunogen sequence of anti-ARF6 antibody PB9987 to see if they may cross-react. If the sequence homology is close, then you can perform a pilot test. Keep in mind that since we have not validated canine samples, this use of the antibody is not covered by our guarantee. However we have an innovator award program that if you test this antibody and show it works in canine leukocyte in Flow Cytometry, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-06-05
Question
I see that the anti-ARF6 antibody PB9987 works with WB, what is the protocol used to produce the result images on the product page?
T. Singh
Verified customer
Asked: 2019-05-23
Answer
You can find protocols for WB 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: 2019-05-23
Question
We have seen staining in rat trachea. Are there any suggestions? Is anti-ARF6 antibody supposed to stain trachea positively?
Verified Customer
Verified customer
Asked: 2018-07-10
Answer
From what I have seen in literature trachea does express ARF6. From what I have seen in Uniprot.org, ARF6 is expressed in esophagus squamous epithelium, umbilical cord blood, brain, trachea, skin uterus, leukocyte, liver, among other tissues. Regarding which tissues have ARF6 expression, here are a few articles citing expression in various tissues:
Leukocyte, Pubmed ID: 11062263
Liver, Pubmed ID: 24275569
Skin, and Uterus, Pubmed ID: 15489334
Trachea, Pubmed ID: 14702039
Umbilical cord blood, Pubmed ID: 9653160
Boster Scientific Support
Answered: 2018-07-10
Question
Will anti-ARF6 antibody PB9987 work for WB with liver?
Verified Customer
Verified customer
Asked: 2017-10-25
Answer
According to the expression profile of liver, ARF6 is highly expressed in liver. So, it is likely that anti-ARF6 antibody PB9987 will work for WB with liver.
Boster Scientific Support
Answered: 2017-10-25
Question
Would PB9987 anti-ARF6 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
E. Patel
Verified customer
Asked: 2017-10-23
Answer
As indicated on the product datasheet, PB9987 anti-ARF6 antibody as been validated on WB. 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: 2017-10-23
Question
I have attached the WB image, lot number and protocol we used for liver using anti-ARF6 antibody PB9987. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2017-07-27
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: 2017-07-27
Question
Can you help my question with product PB9987, anti-ARF6 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: 2017-07-14
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 PB9987 anti-ARF6 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: 2017-07-14
Question
My lab would like using your anti-ARF6 antibody for negative regulation of receptor-mediated endocytosis studies. Has this antibody been tested with western blotting on rat kidney tissue? We would like to see some validation images before ordering.
K. Moore
Verified customer
Asked: 2016-08-01
Answer
We appreciate your inquiry. This PB9987 anti-ARF6 antibody is tested on rat kidney tissue, mouse kidney tissue, siha cells. It is guaranteed to work for Flow Cytometry, 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: 2016-08-01
Question
Is this PB9987 anti-ARF6 antibody reactive to the isotypes of ARF6?
G. Bhatt
Verified customer
Asked: 2014-05-08
Answer
The immunogen of PB9987 anti-ARF6 antibody is E.coli-derived human ARF6 recombinant protein (Position: G2-S175). Human ARF6 shares 100% amino acid (aa) sequence identity with both mouse and rat ARF6. 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: 2014-05-08
Question
Is a blocking peptide available for product anti-ARF6 antibody (PB9987)?
C. Wu
Verified customer
Asked: 2013-11-25
Answer
We do provide the blocking peptide for product anti-ARF6 antibody (PB9987). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2013-11-25