Anti-Clathrin antibody [X22] - Membrane Vesicle Marker (ab2731)

infos sur des marqueurs fluo des voies d'internalisation.

ANSWER:

Nous avons dans notre catalogue des anticorps marqueurs des voies d'internalisation tels que les anti-Clathrin ab2731 (http://www.abcam.com/ab2731) et ab59710 (http://www.abcam.com/ab59710). Malheureusement nous n'avons pas ces anticorps couplés à un fluorochrome.

Nous avons cependant des kits de conjugaison, les kits EasyLink, qui permettent de coupler rapidement et facilement les anticorps primaire. Pour plus d'informations sur ces kits je vous invite à visiter la page suivante : http://www.abcam.com/easylink.

Pour les voies d'internalisation je vous suggère le lien suivant : http://www.abcam.com/index.html?pageconfig=resource&rid=10236&pid=14.

J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

thank you for your reply.

Our customer would like to know if you have any abreview for both or for just one these batches about WB analysis.

If so, please send me all the information you have.

Thanks in advance

Kind regards

ANSWER:

Thank you for your reply.

All Abreviews we have are from previous batches of this antibody. This antibody is very popular and sells fast. Therefore even the experiment shown in the latest Abreview was performed with a different batch.

I am sorry I cannot provide you with this information, but I would like to reassure you and your customer that at Abcam our products are covered by the Abpromise.

Our Abpromise® ensures that you can trust our products to perform as stated on the datasheets, or we will offer a replacement, credit, or refund.

The Abcam Abpromise® guarantee:

• 100% Scientific and Customer Support for any product you buy from Abcam or one of our authorised distributors.

• We guarantee our products work in the tested species and applications stated on the datasheet.

• We will replace or refund products not performing as stated on the datasheet if reported within 6 months of purchase.

• We investigate any quality concerns raised by customers to ensure our catalog contains products that perform to the highest standards.

Please note these conditions to our Abpromise®:

• Protocol information must be provided in order for the claim to be validated.

• Antibodies tested in recombinant samples only are not guaranteed for use on endogenous samples.

• “Predicted to react” information on the datasheets is provided for reference only and these species are not guaranteed.

• Fast Track antibodies are covered for ELISA against the immunizing peptide only.

I hope this information is helpful, but please do not hesitate to contact us with further questions.

I have been having some problems with this new lot85960, I have run to experiments and I do not see any bands, I used what you recomend for western blots 1:500. I used lot73284 and was OK.

ANSWER:

I'm very sorry to hear you are having problems with your recent vial of ab2731. Having checked the lot numbers I was able to find out the two vials came from the same master stock and we have not received other complaints regarding this lot so it is likely that the vial was damaged during shipping. I have arrange for a replacement vial to be sent to you, you should receive a confirmation e-mail soon (order 76724).

Please let me know if you need further assistance and I will do my best to help you,

DESCRIPTION OF THE PROBLEM Multiple bands. Very weak band at 180 kDa, and stronger bands elsewhere, f.ex. at around 100 and 80 kDa.

SAMPLE Lysate of HeLa cells.

PRIMARY ANTIBODY ab2731 anti-Clathrin was diluted 1:500 or 1:1000 in TBS (10 mM Tris/HCl (pH 7.4)/137 mM NaCl] with 0.1% (v/v) Tween 20) containing 1% fat-free dry-milk. Blots were incubated overnight at 4?C, before washing 3x 10 min in TBS, and then blocking for 30 min.

SECONDARY ANTIBODY Peroxidase-conjugated Donkey anti-Mouse IgG from Jackson ImmunoResearch diluted 1:4000 in TBS containing 1% fat-free dry-milk. Incubated 2h at room temperature, before washing 3x 10 min in TBS.

DETECTION METHOD ECL

POSITIVE AND NEGATIVE CONTROLS USED HeLa lysate should have plenty of Clathrin. No negative control.

ANTIBODY STORAGE CONDITIONS 4?C

SAMPLE PREPARATION Cells were lysed in SDS/PAGE sample buffer [10 mM Tris/HCl (pH 6.8)/5 mM EDTA/50 mM NaF/30 mM sodium pyrophosphate/2% (w/v) SDS/1 mM Na3VO4/1 mM PMSF] on ice for 10 min. Then 4% (v/v) glycerol, 4% (v/v) b-mercaptoethanol and 0.005% (w/v) Bromophenol Blue were added to the lysates. The lysates were incubated at 95?C for 10 min.

AMOUNT OF PROTEIN LOADED Sufficient. Normal sample amount.

ELECTROPHORESIS/GEL CONDITIONS Reducing 10 % gel run at 150 V for 1-1,5 hours in standard electrophoresis buffer

TRANSFER AND BLOCKING CONDITIONS Transferred to Nitrocellulose membrane at 100 V for 1 hour in standard transfer buffer. Blocking solution Tris-buffered saline [TBS; 10 mM Tris/HCl (pH 7.4)/137 mM NaCl] with 0.1% (v/v) Tween 20, containing 5% fat-free dry-milk. Membrane was blocked for 30 min at room temp.

ANSWER:

Thank you for your enquiry and for forwarding some more information about your Western blot assay.

Looking through your protocol, we notice that you stored the antibody at 4oC. This may be only part of the problem, we recommend -20oC storage.

The other advice would be to use fresh proteinase inhibitors in order to inhibit protein degradation.

Should you still have problem with this antibody, then please do not hesitate to get back to us.

We hope this will help.

Hello: Please can you tell me if you every offer free evaluation samples of antibodies for customers to try in their own applications? I have a customer who is intersted in the mouse antibody against clathrin (cat. # ab2731) and who is interested in trying the product with HUBEC cells for localization of clathrin in conjunction with potassium channels. If you could offer a small sample, she would be very grateful. I look forward to hearing from you. Thank you for your help. Best Regards, Liz

ANSWER:

We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement.

Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will forward a GBP10/ USD15/ EUR15 Amazon gift voucher.