Overview

  • Product name
  • Description
    Rabbit polyclonal to ATG7
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-FoFr, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Chicken, Human
  • Immunogen

    Synthetic peptide corresponding to Human ATG7 (C terminal).

  • Positive control
    • Caco-2 cell lysate, MCF7 cells.

Applications

Our Abpromise guarantee covers the use of ab53255 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-FoFr Use at an assay dependent concentration. PubMed: 21784844
ICC/IF Use a concentration of 5 - 10 µg/ml.
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 78 kDa (predicted molecular weight: 78 kDa).
IHC-P Use at an assay dependent concentration. PubMed: 20174468

Target

  • Function
    E1-like activating enzyme involved in the 2 ubiquitin-like systems required for cytoplasm to vacuole transport (Cvt) and autophagy. Activates ATG12 for its conjugation with ATG5 as well as the ATG8 family proteins for their conjugation with phosphatidylethanolamine. Both systems are needed for the ATG8 association to Cvt vesicles and autophagosomes membranes. Required for autophagic death induced by caspase-8 inhibition. Required for mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Modulates p53/TP53 activity to regulate cell cycle and survival during metabolic stress. Plays also a key role in the maintenance of axonal homeostasis, the prevention of axonal degeneration, the maintenance of hematopoietic stem cells, the formation of Paneth cell granules, as well as in adipose differentiation.
  • Tissue specificity
    Widely expressed, especially in kidney, liver, lymph nodes and bone marrow.
  • Sequence similarities
    Belongs to the ATG7 family.
  • Domain
    The C-terminal part of the protein is essential for the dimerization and interaction with ATG3 and ATG12.
    The N-terminal FAP motif (residues 15 to 17) is essential for the formation of the ATG89-PE and ATG5-ATG12 conjugates.
  • Post-translational
    modifications
    Acetylated by EP300.
  • Cellular localization
    Cytoplasm. Preautophagosomal structure. Localizes also to discrete punctae along the ciliary axoneme and to the base of the ciliary axoneme.
  • Information by UniProt
  • Database links
  • Alternative names
    • 1810013K23Rik antibody
    • Apg 7 antibody
    • APG7 autophagy 7 like antibody
    • APG7 autophagy 7-like (S. cerevisiae) antibody
    • APG7 like antibody
    • APG7, S. cerevisiae, homolog of antibody
    • APG7-like antibody
    • APG7L antibody
    • ATG 7 antibody
    • ATG12-activating enzyme E1 ATG7 antibody
    • ATG7 antibody
    • ATG7 autophagy related 7 homolog (S. cerevisiae) antibody
    • ATG7 autophagy related 7 homolog antibody
    • ATG7_HUMAN antibody
    • Atg7l antibody
    • Autophagy 7, S. cerevisiae, homolog of antibody
    • Autophagy related protein 7 antibody
    • Autophagy-related 7 (yeast) antibody
    • Autophagy-related protein 7 antibody
    • DKFZp434N0735 antibody
    • GSA 7 antibody
    • GSA7 antibody
    • hAGP7 antibody
    • Ubiquitin activating enzyme E1 like protein antibody
    • Ubiquitin-activating enzyme E1-like protein antibody
    • Ubiquitin-like modifier-activating enzyme ATG7 antibody
    see all

Images

  • Lane 1 : Anti-ATG7 antibody (ab53255) at 0.5 µg/ml
    Lane 2 : Anti-ATG7 antibody (ab53255) at 1 µg/ml
    Lane 3 : Anti-ATG7 antibody (ab53255) at 2 µg/ml

    All lanes : Caco-2 cell lysate at 15 µg per lane.

    Predicted band size: 78 kDa
    Observed band size: 78 kDa

  • This image shows ab53255 staining MCF7 cells as 10µg/ml
  • IHC image of ab53255 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab53255, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemical analysis of murine brain tissue, staining ATG7 with ab53255. Fixed brain sections were blocked with serum, permeabilized with 0.2% Triton X-100, and treated with primary antibody. An AlexaFluor®555-conjugated anti-rabbit IgG was used as the secondary antibody.

References

This product has been referenced in:
  • Chen L  et al. Juglanin inhibits lung cancer by regulation of apoptosis, ROS and autophagy induction. Oncotarget 8:93878-93898 (2017). Read more (PubMed: 29212196) »
  • Sun S  et al. ATG7 promotes the tumorigenesis of lung cancer but might be dispensable for prognosis predication: a clinicopathologic study. Onco Targets Ther 9:4975-81 (2016). IHC-P ; Human . Read more (PubMed: 27563251) »
See all 19 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Sample
Chicken Cell (Retinal Ganglion Cell culture)
Specification
Retinal Ganglion Cell culture
Permeabilization
Yes - PBS + 0.1% Tween20
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 10 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Sample
Mouse Tissue sections (Uterus)
Specification
Uterus
Permeabilization
No
Fixative
Paraformaldehyde

Dr. Mukesh Jaiswal

Verified customer

Submitted Nov 13 2013

Question
Answer

Thank you for confirming the PO number and for having provided us with your protocol.The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement (ab133528) with the order number xxxxxxx.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

Thank you for contacting us and for providing us with extra information about the protocol used.

All our products are covered by the Abpromise guarentee and since our product did not work as stated on the datasheet, I would be happy to replace the antibody ab53255 with ab133528 providing the product was purchased in the last six months.

In order for me to issue this free of charge replacement, I need the orginal order number of ab53255 and I can't seem to find it in our records. Was this product purchased in the last 6 months and if it was, do you have the orginal order number so that I may issue you a free of charge replacement?

I look forward to receiving your reply.

Read More

Answer

Thank you for taking time to contact us.

I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. Could you please additionally tell us the blocking agent used as this will help us further investigate this product?

I can recommend two other antibodies which are guaranteed to work in Western blot in mouse: ab133528 (https://www.abcam.com/ab133528) or ab80639 (https://www.abcam.com/ab80639). Alternatively I could give you a refund for this antibody.

Once again I apologize for the inconvenience and if you could let me know which resolution you would prefer, I would be pleased to offer you a free of charge replacement, credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (THP-1 monocyte/macrophage cell line)
Specification
THP-1 monocyte/macrophage cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Aug 07 2012

Answer

Thank you very much for your interest in ab53255.

To our knowledge, ab53255 has not been tested in FACS. I can recommend ab52472 which has been tested if you are using human samples. It does not react with mouse or rat. Alternatively, I can offer a discount off a future purchase if you buy ab53255 now, test it in FACS and submit feedback to us in the form of an Abreview. It doesn't matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab53255 in FACS. I will then send a discount code. This code must be issued before purchasing ab53255 so please wait for my reply before ordering.

2. Purchase ab53255 either by phone, fax, or online (www.abcam.com).

3. Test it in FACS.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody] ordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab53255 turns out to be unsuitable for FACS, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for contacting us.

Your credit note ID is 20XXX (for 1 vial of ab91526).

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Read More

Question

thank you for your prompt answer. Here below the answers to your questions. I hope this could help in solving the problem.

1. Do you know the lot number and order number of each of the antibodies?
The lot numbers are the following: Apg7 Lot GR54674-4

p62 GR7582-8

While for the order number at the moment I can’t provide you any information as the technician who ordered the antibodies for me is now on holiays.

2. How was the sample collected and treated prior to loading on the gel? What lysis buffer was used? Were any protease inhibitors used? Was the sample boiled prior to loading? Was any reducing agent used?
Samples were first stored at -80 (after dissection of the animal), thawed in ice. Lysis buffer added and samply lysed by sonication with two pulses for 5 sec.
Lysis buffer: SDS 1% (or triton X 100 1% in a second experiment), EDTA 1mM, NaCl 50mM and TrisHcl pH 7.4 100mM.
Complete protease and phospatase inhibitors were freshly added to the lysate.
Samples were boiled for 10min at 95C after adding loading buffer.
No reducing agents were used.

3. How was the blocking performed? Time/temperature? Was any detergent included in the blocking buffer?
Blocking was done in different blocking buffers: milk 5% or BSA 5% or NGS 5%. 1h at room temperature. Blocking buffers were diluted in TBS + 0.1% Tween 20.

4. How was the primary antibody incubated with the membrane (time/temperature)? What diluent was used with each of the antibodies?
Antibodies were incubated O/N at 4C. Antibodies were diluted in milk 2%, BSA 5% or NGS 5% in TBS + 0.1% Tween 20.

5. What washing steps were employed and with what buffer?
Washing steps: 3x 10 min in TBS + 0.1% Tween 20.

6. Were the membranes freshly probed or was the same membrane used and stripped between each antibody? If so, how was this procedure performed?
Membranes were all freshly probed.

7. Secondary antibody:goat anti rabbit, used 1:10’000 in milk 2%, BSA 5% or NGS 5% in TBS + 0.1% Tween 20. 1h at room temperature.

All the best,

Read More
Answer

Thank you for providing that information.

Having reviewed the protocol used, I believe there are afew things that maybe worthtrying in order toimprove the specificity observed.I would suggest using a reducing agent in the loading buffer to reduce the aggregation which may be occurring (especially with ab91526). Either DTT or b-mecaptoethanol. It would also be worth reducing the temperature at which the protein is denatured. Certain proteins are prone to aggregation, which can be reduced by heating to 70 degrees for 10 minutes instead of 95. I would also suggest reducing the concentration used slightly (ab53255 down to 1/500 and ab91526 to 1/1000) to see if this improves the binding as well as reducing the amount of protein loaded (down to 40 ug/lane).

Has a "no primary" control been performed? This may be worthwhile, just to rule out any non-specificity relating to the secondary antibody used.

Should these suggestions not improve the results please do let me know and I'll look into if we have any alternative lots of these antibodiesor different antibodies altogetherwhich you may be interested in trying.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewedthe information you have provided, I would like to confirm a few further details in order to understand the procedure used more fully and hopefully provide more specific protocol tips:

1. Do you know the lot number and order number of each of the antibodies?

2. How was the sample collected and treated prior to loading on the gel? What lysis buffer was used? Were any protease inhibitors used? Was the sample boiled prior to loading? Was any reducing agent used?

3. How was the blocking performed? Time/temperature? Was any detergent included in the blocking buffer?

4. How was the primary antibody incubated with the membrane (time/temperature)? What diluent was used with each of the antibodies?

5. What washing steps were employed and with what buffer?

6. Were the membranes freshly probed or was the same membrane used and stripped between each antibody? If so, how was this procedure performed?

Hopefully with this information I will be ableto suggest modificationswhich will improve the specificity currently observed.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I look forward to receiving your reply.

Read More

1-10 of 13 Abreviews or Q&A

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