Overview

  • Product name

  • Description

    Rabbit polyclonal to Dock5
  • Host species

    Rabbit
  • Specificity

    This antibody reacts with 23 & 215 kDa mouse proteins and the 215 kDa human protein.
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide derived from the C terminal domain of mouse Dock5 protein

Properties

Applications

Our Abpromise guarantee covers the use of ab71578 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
WB 1/200 - 1/2000. Predicted molecular weight: 215 kDa.

Target

References

ab71578 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Answer

Thank you for contacting us. Your credit note ID is xxxx. I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. 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.

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Answer

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments, and would like to offer a refund or credit note in compensation as you had contacted us within the 6 month Abpromise guarantee period. In order to arrange this, I would appreciate if you could confirm your order number and date of purchase?

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

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Question
Answer

Thank you for participating in our satisfaction survey.

I do look forward to hearing from you as to whether this product's performance has improved but am troubled to hear that you as experiencing issues with ab76402. Please feel free to contact me regarding these products as I would very much like to help you to resolve these issues.

Please remember that our products are covered by Abpromise guarantee. Should a product not perform as indicated on the datasheet, we gladly provide scientific support, replacement or refund of that product. More information on our Abpromise may be found at the following link:

https://www.abcam.com/index.html?pageconfig=abpromise

I sympathize with your frustration when it comes to import restrictions to Canada. However while this might limit the availability of some products to you, our goal is to have every Abcam product which you purchase give you the best results.

Please do not hesitate contact me if you have any questions now or in the future.

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Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (Human HaCaT keratinocytes)
Loading amount
30 µg
Specification
Human HaCaT keratinocytes
Gel Running Conditions
Reduced Denaturing (7.5%)
Blocking step
LiCor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C

Dr. Leeni Koivisto

Verified customer

Submitted Jun 20 2012

Answer

Thank you for submitting an Abreview for Ab71578.

We were surprised to see that you were unable to get satisfactory results when using this product in western blot on human cell types. As this product is guaranteed to work in this application, I would like offer assistance to help you to achieve the best results possible. The protocol details which you have provided us indicate that this product should have worked however I was wondering if you could provide me with some additional information. Could you outline any troubleshooting steps which you attempted such as changes in antibody or protein concentrations, transfer procedures, positive or negative controls and the like?


This product is covered by our Abpromise guarantee. If we cannot remedy this issue and this is a product that you have purchased within the last six months, we will replace or refund it under our Abpromise guarantee, as you are using it according to specifications listed on our datasheet.
I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for your response.

While there were non specific bands lower in the gel which are around 50kDA, the upper region of interest appeared very near the top directly below the stacking layer were there seemed to be a wavy band. Running the gel longer will allow any proteins which may be in this area to resolve. As there is no non specific binding in this region use of the higher concentration of antibody will not have adverse affect and may approach the threshold for affinity of the product with the target.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

Hello, We purchased the rabbit anti-human DOCK5 antibody ab71578 in January 2012 to visualize the strong signals for this protein that we have obtained from our mass spectrometry analyses by Western blotting. Since you didn't have a picture of a WB included on your product info, I agreed to do an Abreview for this antibody with the image for WB.   Before I submit my Abreview and ask for a refund for this antibody, I'm wondering whether you would like to comment on these WBs and maybe troubleshoot the issues with me.   Details of the experiments with pictures in the attached file:   Panel A. Human keratinocyte lysates were immunoprecipitated with anti-human GSK3a antibody cross-linked to magnetic Dynabeads . Control IP was performed using the respective non-immune IgG. Co-immunoprecipitated complexes  eluted in glycine-HCl pH 2.8 and separated in a 4-15% gradient gel. Proteins were visualized by silver staining. Panel A. The gel was sliced, and the proteins within each of these fractions were analysed by mass spectrometry. The major protein specifically recognized in the fraction 2 in the sample immunoprecipitated with the GSK3a antibody was identified as human DOCK5. (The only other protein recognized was human keratin type II cytoskeletal, but this was also detected in the negative control sample.)    We then purchased the above-mentioned DOCK5 antibody to verify these results by WB and to further study the protein interactions. The pictures (C-E) show WBs from whole cell lysates (reduced samples) separated in 4/7.5% SDS-PAGE gels.  The general protocol was: The sample was run with  a molecular weight marker (Bio-Rad Precision Plus All Blue; 1 µl; shows up mainly in the 700 channel of the infrared reader) and transferred onto Hybond-ECL nitrocellulose membrane (Amersham, GE Healtcare). The membranes were blocked by incubating in Odyssey Blocking Buffer (Li-Cor) for 1h at RT. The primary antibody were diluted in the same blocking solution containing 0.1% Tween 20 and added to the membranes for o/n at +4ºC. The membranes were washed twice with TBST and then incubated with appropriate IRDye secondary antibodies (Li-Cor)  diluted 1/20,000 in the same blocking solution containing 0.1% Tween 20 and 0.02% SDS for 1h at RT. Membranes were then washed twice briefly and then four times 5min with TBST, followed by a 5min wash with TBS.  The detection was performed using Li-Cor Odyssey infrared reader.  Panel C. Primary antibody dilution 1/1000 (Abcam recommended dilution range for WB: 1/200 - 1/2000), secondary antibody used was donkey anti-rabbit IRDye800. Panel D. To exclude the possibility that the background results from the secondary antibody, the blot was redone using goat anti-rabbit IRDye680 as the secondary antibody. The signal obtained with this secondary antibody shows on a detection channel that is different from IRDye800)  Panel E. Another gel probed with the primary antibody further diluted to 1/2000 followed by goat anti-rabbit IRDye680   I was not happy with the results of my Western blots. The antibody was  non-specific, resulting in numerous bands in the blots. Moreover, it didn't recognize a protein with the indicated molecular weight (215 kDa). There is a faint band at >250 kDa, but it is not matching the protein seen in panels A and B.   Diluting the antibodies further or increasing the washing times did not improve the  specificity of antibody binding.    Looking forward for hearing back from you.

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Answer

Thank you for contacting us.

I am sorry to hear of the troubles which you have been experiencing with this product. Having reviewed the protocol and images which you have kindly provided I would like to suggest a few suggestions which may help.

DOCK5 should be over 200kDA when transferring large proteins we suggest adding SDS to a final concentration of 0.1% in the transfer buffer and reducing the methanol percentage to 10%. Methanol is only necessary when using nitrocellulose membranes and may be removed all together when using PVDF.

I would also suggest running your sample in a low concentration gel, 8% or less and allowing the sample to migrate farther.

Finally I would suggest using lower dilutions of this antibody such as 1:500 or 1:200.

Your Abreview code will be redeemable regardless of positive or negative review. Please be sure to include the code in the additional notes section of your review.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.


Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

I am very pleased to hear you would like to use ab71578 in WB and send us an image of your results. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview with the image for WB and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. The code will be active once the Abreview with the image has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated. Remember if the product does not work you will be covered by our Abpromise guarantee and eligible for a full refund or replacement. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount

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Answer

Thank you very much for your interest in ab71578. Unfortunately we do not currently have a WB image for ab71578, however it has been tested in and we do guarantee it for WB. Therefore, I can offer a discount off a future purchase if you buy ab71578 now, use it WB and submit an image of your results to us in the form of an Abreview. The discount would be to the value of 1 free primary antibody. It doesn’t matter whether the result is positive or negative, we would just really like to receive your feedback. Of course, if the results are negative you will be covered by our Abpromise guarantee and eligible for a refund or replacement. 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 send an image of ab71578 in WB. I will then send a discount code. This code must be issued before purchasing ab71578 so please wait for my reply before ordering. 2. Purchase ab71578 either by phone, fax, or online (www.abcam.com). 3. Use it in WB. 4. Send us an image of the results, positive or negative, using our Abreview system (this will take about 10 minutes). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review with the image 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. We are always pleased to obtain feedback about our products and any information is greatly appreciated! 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.

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