Product Info Summary
SKU: | PB9481 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-ATG14L Antibody Picoband™
SKU/Catalog Number
PB9481
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ATG14L Antibody Picoband™ catalog # PB9481. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, 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-ATG14L Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9481)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the N-terminus of human ATG14L, different from the related mouse and rat sequences by two amino acids.
*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
PB9481 is reactive to ATG14 in Human, Rat
Applications
PB9481 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
59 kDa
Calculated molecular weight
55.309kDa
Background of ATG14
ATG14 (also known as beclin-1-associated autophagy-related key regulator (Barkor) or ATG14L), an essential autophagy-specific regulator of the class III phosphatidylinositol 3-kinase complex, promotes membrane tethering of protein-free liposomes, and enhances hemifusion and full fusion of proteoliposomes reconstituted with the target (t)-SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) syntaxin 17 (STX17) and SNAP29, and the vesicle (v)-SNARE VAMP8 (vesicle-associated membrane protein 8). ATG14 binds to the SNARE core domain of STX17 through its coiled-coil domain, and stabilizes the STX17-SNAP29 binary t-SNARE complex on autophagosomes.
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.
Innovating Scientists Reward
If you are the first to review this product, or if you have results for a special sample, species or application this product is not validated in, share your results with us and receive product credits you can use towards any Boster products! Applicable to all scientists worldwide.
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Assay dilution & Images
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, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Immunofluorescence, 5 μg/ml, Human, Rat
Flow Cytometry, 1-3μg/1x106 cells, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of ATG14L using anti-ATG14L antibody (PB9481).
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 Brain Tissue Lysate,
Lane 2: HELA Whole Cell Lysate.
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-ATG14L antigen affinity purified polyclonal antibody (Catalog # PB9481) 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 ATG14L at approximately 59KD. The expected band size for ATG14L is at 59KD.
Click image to see more details
Figure 2. IHC analysis of ATG14L using anti-ATG14L antibody (PB9481).
ATG14L was detected in paraffin-embedded section of Rat Spleen Tissue. 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-ATG14L Antibody (PB9481) 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 3. IHC analysis of ATG14L using anti-ATG14L antibody (PB9481).
ATG14L was detected in paraffin-embedded section of Human Lung Cancer Tissue. 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-ATG14L Antibody (PB9481) 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. IF analysis of ATG14L using anti-ATG14L antibody (PB9481).
ATG14L was detected in an immunocytochemical section of U2OS 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 5 μg/mL rabbit anti-ATG14L Antibody (PB9481) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 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.
Click image to see more details
Figure 5. IF analysis of ATG14L using anti-ATG14L antibody (PB9481).
ATG14L was detected in a paraffin-embedded section of human lung 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 5 μg/mL rabbit anti-ATG14L Antibody (PB9481) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 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.
Click image to see more details
Figure 6. IF analysis of ATG14L using anti-ATG14L antibody (PB9481).
ATG14L was detected in a paraffin-embedded section of human colon 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 5 μg/mL rabbit anti-ATG14L Antibody (PB9481) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 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.
Click image to see more details
Figure 7. IF analysis of ATG14L using anti-ATG14L antibody (PB9481).
ATG14L was detected in a paraffin-embedded section of rat spleen 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 5 μg/mL rabbit anti-ATG14L Antibody (PB9481) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 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.
Click image to see more details
Figure 8. Flow Cytometry analysis of SiHa cells using anti-ATG14L antibody (PB9481).
Overlay histogram showing SiHa cells stained with PB9481 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG14L Antibody (PB9481,1μg/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 (Red line) was also used as a control.
Click image to see more details
Figure 9. Flow Cytometry analysis of A431 cells using anti-ATG14L antibody (PB9481).
Overlay histogram showing A431 cells stained with PB9481 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG14L Antibody (PB9481,1μg/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 (Red line) was also used as a control.
Click image to see more details
Figure 10. Flow Cytometry analysis of PC-3 cells using anti-ATG14L antibody (PB9481).
Overlay histogram showing PC-3 cells stained with PB9481 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG14L Antibody (PB9481,1μg/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 (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For ATG14 (Source: Uniprot.org, NCBI)
Gene Name
ATG14
Full Name
Beclin 1-associated autophagy-related key regulator
Weight
55.309kDa
Superfamily
ATG14 family
Alternative Names
ATG14 Autophagy Related 14 Homolog (S. Cerevisiae); ATG14 Autophagy Related 14 Homolog; ATG14L; Autophagy Related 14; Autophagy-Related Protein 14-Like Protein; Barkor; Beclin 1-Associated Autophagy-Related Key Regulator; Beclin 1-interacting protein; KIAA0831 ATG14 ATG14L, BARKOR, KIAA0831 autophagy related 14 beclin 1-associated autophagy-related key regulator|ATG14 autophagy related 14 homolog|Beclin 1-Interacting protein|autophagy-related protein 14-like protein
*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 ATG14, check out the ATG14 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 ATG14: 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-ATG14L Antibody Picoband™ (PB9481)
Hello CJ!
PB9481 has been cited in 1 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.
Polydatin induces apoptosis and autophagy via STAT3 signaling in human osteosarcoma MG-63 cells
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Customer Reviews
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Customer Q&As
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17 Customer Q&As for Anti-ATG14L Antibody Picoband™
Question
My lab would like to test anti-ATG14L?Picoband™ antibody PB9481 on human female gonad for research purposes, then I may be interested in using anti-ATG14L?Picoband™ antibody PB9481 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2020-04-27
Answer
The products we sell, including anti-ATG14L?Picoband™ antibody PB9481, 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: 2020-04-27
Question
Would anti-ATG14L?Picoband™ antibody PB9481 work on dog Flow Cytometry with cervix carcinoma?
Verified Customer
Verified customer
Asked: 2020-04-24
Answer
Our lab technicians have not validated anti-ATG14L?Picoband™ antibody PB9481 on dog. You can run a BLAST between dog and the immunogen sequence of anti-ATG14L?Picoband™ antibody PB9481 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 dog 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 dog cervix carcinoma in Flow Cytometry, you can get your next antibody for free.
Boster Scientific Support
Answered: 2020-04-24
Question
I was wanting to use your anti-ATG14L?Picoband™ antibody for ICC for human female gonad 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 female gonad identification?
Verified Customer
Verified customer
Asked: 2020-02-14
Answer
As indicated on the product datasheet, PB9481 anti-ATG14L?Picoband™ antibody has been tested for Flow Cytometry, IHC, ICC, WB on human, rat tissues. We have an innovator award program that if you test this antibody and show it works in human female gonad in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2020-02-14
Question
Is a blocking peptide available for product anti-ATG14L?Picoband™ antibody (PB9481)?
Verified Customer
Verified customer
Asked: 2019-11-29
Answer
We do provide the blocking peptide for product anti-ATG14L?Picoband™ antibody (PB9481). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2019-11-29
Question
We are currently using anti-ATG14L?Picoband™ antibody PB9481 for rat tissue, and we are well pleased with the ICC results. The species of reactivity given in the datasheet says human, rat. Is it possible that the antibody can work on bovine tissues as well?
Verified Customer
Verified customer
Asked: 2019-10-14
Answer
The anti-ATG14L?Picoband™ antibody (PB9481) has not been tested for cross reactivity specifically with bovine tissues, though 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 bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-10-14
Question
We want using your anti-ATG14L?antibody for cellular response to starvation studies. Has this antibody been tested with western blotting on siha cells? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2019-06-17
Answer
We appreciate your inquiry. This PB9481 anti-ATG14L?antibody is validated on rat spleen tissue, brain tissue, tissue lysate, hela whole cell lysate, lung cancer tissue, siha cells, a431 cells. It is guaranteed to work for Flow Cytometry, IHC, ICC, WB in human, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2019-06-17
Question
Please see the WB image, lot number and protocol we used for female gonad using anti-ATG14L?Picoband™ antibody PB9481. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2019-05-09
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: 2019-05-09
Question
My team were well pleased with the WB result of your anti-ATG14L?Picoband™ antibody. However we have seen positive staining in hippocampus trachea cytoplasm. using this antibody. Is that expected? Could you tell me where is ATG14 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-02-28
Answer
From what I have seen in literature, hippocampus trachea does express ATG14. Generally ATG14 expresses in cytoplasm. Regarding which tissues have ATG14 expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 10048485
Cervix carcinoma, Pubmed ID: 17081983
Hippocampus, and Trachea, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2019-02-28
Question
Thank you for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for female gonad using anti-ATG14L?Picoband™ antibody PB9481. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2018-05-28
Answer
I appreciate 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: 2018-05-28
Question
We have observed staining in rat brain. What should we do? Is anti-ATG14L?Picoband™ antibody supposed to stain brain positively?
Verified Customer
Verified customer
Asked: 2018-04-09
Answer
Based on literature brain does express ATG14. Based on Uniprot.org, ATG14 is expressed in female gonad, brain, hippocampus trachea, cervix carcinoma, among other tissues. Regarding which tissues have ATG14 expression, here are a few articles citing expression in various tissues:
Brain, Pubmed ID: 10048485
Cervix carcinoma, Pubmed ID: 17081983
Hippocampus, and Trachea, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2018-04-09
Question
I see that the anti-ATG14L?Picoband™ antibody PB9481 works with ICC, what is the protocol used to produce the result images on the product page?
J. Johnson
Verified customer
Asked: 2017-12-14
Answer
You can find protocols for ICC 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: 2017-12-14
Question
Would PB9481 anti-ATG14L?Picoband™ antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2017-11-16
Answer
As indicated on the product datasheet, PB9481 anti-ATG14L?Picoband™ antibody as been tested on ICC. 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: 2017-11-16
Question
Our lab used your anti-ATG14L?Picoband™ antibody for Flow Cytometry on hippocampus trachea last year. I am using rat, and We want to use the antibody for IHC next. I am interested in examining hippocampus trachea as well as brain in our next experiment. Could give a recommendation on which antibody would work the best for IHC?
J. Parker
Verified customer
Asked: 2016-03-21
Answer
I looked at the website and datasheets of our anti-ATG14L?Picoband™ antibody and it appears that PB9481 has been tested on rat in both Flow Cytometry and IHC. Thus PB9481 should work for your application. Our Boster satisfaction guarantee will cover this product for IHC in rat even if the specific tissue type has not been validated. We do have a comprehensive range of products for IHC detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2016-03-21
Question
I have a question about product PB9481, anti-ATG14L?Picoband™ 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?
C. Evans
Verified customer
Asked: 2015-08-24
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 PB9481 anti-ATG14L?Picoband™ 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: 2015-08-24
Question
Does anti-ATG14L?Picoband™ antibody PB9481 work for ICC with female gonad?
B. Parker
Verified customer
Asked: 2014-01-24
Answer
According to the expression profile of female gonad, ATG14 is highly expressed in female gonad. So, it is likely that anti-ATG14L?Picoband™ antibody PB9481 will work for ICC with female gonad.
Boster Scientific Support
Answered: 2014-01-24
Question
Do you have a BSA free version of anti-ATG14L?Picoband™ antibody PB9481 available?
B. Gonzalez
Verified customer
Asked: 2013-12-03
Answer
I appreciate your recent telephone inquiry. I can confirm that some lots of this anti-ATG14L?Picoband™ antibody PB9481 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: 2013-12-03
Question
Is this PB9481 anti-ATG14L?Picoband™ antibody reactive to the isotypes of ATG14?
A. Jones
Verified customer
Asked: 2013-05-30
Answer
The immunogen of PB9481 anti-ATG14L?Picoband™ antibody is A synthetic peptide corresponding to a sequence at the N-terminus of human ATG14L (70-101aa RDRERFIDKKERLSRLKSKQEEFQKEVLKAME), different from the related mouse and rat sequences by two amino acids. 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: 2013-05-30