Overview

  • Product name
  • Description
    Mouse monoclonal to Cofilin
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Sheep, Cow, Pig, Cynomolgus monkey
  • Immunogen

    Recombinant full length protein (GST-tag) corresponding to Human Cofilin. AAH11005
    Sequence:

    MASGVAVSDGVIKVFNDMKVRKSSTPEEVKKRKKAVLFCLSEDKKNIILE EGKEILVGDVGQTVDDPYATFAKMLPDKDCRYALYDATYETKESKKEDLV FIFWAPESAPLKSKMIYASSKDAIKKKLTGIKHELQANCYEGVKDRCTLA EKLGGSAVISLEGKPL


    Database link: P23528

  • Positive control
    • WB: HeLa, PC-12 and NIH/3T3 cell lysate. IHC-P: Human breast cancer tissue. ICC/IF: HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab54532 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 Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 18 kDa.
IHC-P Use a concentration of 1.5 µg/ml.
ICC/IF Use a concentration of 10 µg/ml.

Target

  • Function
    Controls reversibly actin polymerization and depolymerization in a pH-sensitive manner. It has the ability to bind G- and F-actin in a 1:1 ratio of cofilin to actin. It is the major component of intranuclear and cytoplasmic actin rods.
  • Tissue specificity
    Widely distributed in various tissues.
  • Sequence similarities
    Belongs to the actin-binding proteins ADF family.
    Contains 1 ADF-H domain.
  • Post-translational
    modifications
    Phosphorylated on Ser-3 in resting cells.
  • Cellular localization
    Nucleus matrix. Cytoplasm > cytoskeleton. Almost completely in nucleus in cells exposed to heat shock or 10% dimethyl sulfoxide.
  • Information by UniProt
  • Database links
  • Alternative names
    • 18 kDa phosphoprotein antibody
    • CFL 1 antibody
    • CFL antibody
    • CFL1 antibody
    • COF1_HUMAN antibody
    • Cofilin 1 antibody
    • Cofilin 1 non muscle antibody
    • Cofilin antibody
    • Cofilin non muscle isoform antibody
    • Cofilin-1 antibody
    • epididymis secretory protein Li 15 antibody
    • HEL-S-15 antibody
    • non-muscle isoform antibody
    • p18 antibody
    see all

Images

  • Anti-Cofilin antibody (ab54532) at 1 µg + HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 25 µg

    Predicted band size: 18 kDa

  • Formalin-fixed, paraffin-embedded human breast cancer tissue stained for Cofilin with ab54532 at 1.5 µg/ml in immunohistochemical analysis.

  • HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for Cofilin (green) using ab54532 at 10 µg/ml in ICC/IF.

  • Anti-Cofilin antibody (ab54532) at 1 µg/ml + NIH/3T3 (mouse embryo fibroblast cell line) cell lysate

    Predicted band size: 18 kDa

  • Anti-Cofilin antibody (ab54532) at 1 µg/ml + PC-12 (rat adrenal gland pheochromocytoma cell line) cell lysate

    Predicted band size: 18 kDa

References

This product has been referenced in:
  • Ye X  et al. Direct dorsal hippocampal-prelimbic cortex connections strengthen fear memories. Nat Neurosci 20:52-61 (2017). Read more (PubMed: 27869801) »
  • Dombert B  et al. BDNF/trkB Induction of Calcium Transients through Cav2.2 Calcium Channels in Motoneurons Corresponds to F-actin Assembly and Growth Cone Formation on ß2-Chain Laminin (221). Front Mol Neurosci 10:346 (2017). Read more (PubMed: 29163025) »
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Rat Cell lysate - whole cell (PC12, rat pheochromacytoma)
Total protein in input
1000 µg
Specification
PC12, rat pheochromacytoma
Immuno-precipitation step
Protein G

Abcam user community

Verified customer

Submitted Oct 31 2012

Answer

Thank you for your reply. I am very pleased to hear you would like to accept our offer and test ab54532 in IP.

Please place your order for ab54532 with the following discount code. This first order will be invoiced at full price:

DISCOUNT CODE: 100%ABR-EB34E
Expiration date: 5th Jan 2013

This code will give you: 1 free primary antibody before the expiration date. To redeem this offer once you have tested the antibody, please submit an Abreview for IP and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.

For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview 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, whether positive or negative. 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.

Read More

Answer

Thank you very much for your reply. I am pleased to hear you are interested in the testing discount offer.

I can offer a discount off a future purchase if you buy ab54532 now, test it in IP 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 ab54532 in IP. I will then send a discount code. This code must be issued before purchasing ab54532 so please wait for my reply before ordering.

2. Purchase ab54532 either by phone, fax, or online (https://www.abcam.com).

I am sorry you will not be able to use the Abpoints for this purchase if you are going to use the testing discount code.

3. Test it in IP.

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 ab 54532 turns out to be unsuitable for IP, 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.

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Answer

Thank you for contacting us and for your interest in our products.

As far as we are aware, ab54532 has never been tested in IP. All tested applications covered by our 6 month guarantee are specified on our datasheets, and these are updated as soon as any new information is brought to our attention.

If you would like to test ab54532Cofilin antibodyin IP, please contact me prior to the purchase by replying to this message as you may be eligible for our testing discount program.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact me for any further advice or information.

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Question

Hi,
I am facing problem with one of your product Ab54532, all detail
is given below,
1) Abcam product code: ab54532
2) Abcam order reference number or product batch number:

3) Description of the problem:
We used this Product for Immunoprecipitation, to capture the
antigen in the cell culture supernatant then analyzed by western blot
by using ab42824 but there was no different between control(only
treated with Magnetic beads) and samples (treated magnetic beads and
ab54532). The Product ab42824 is working very well for normal western
blot. Then we tried Ab54532 for normal western blot by loading 50ug of
protein extract, found that not working from even from 2000-250
dilution. But product Ab42824 is working at 3500 dilution for normal
western blot with 2ug of samples.
4) Sample preparation:
Type of sample (whole cell lysates, fraction, recombinant protein…):
Whole cell lysates
Lysis buffer: Urea lysis buffer
Protease inhibitors: PIC EDTA free
Phosphatase inhibitors
Reducing agent : SDS Sample Buffer
Boiling for ≥5 min? yes
Protein loaded ug/lane or cells/lane: 50ug
Positive control
Negative control
5) Percentage of gel: 12% Acrylamide
Type of membrane: Nitrocellulose
Protein transfer verified: Staining of residual Gel
Blocking agent and concentration: 5% BSA or Milk Powder
Blocking time : 1hr
Blocking temperature: Room Temperature
6) Primary antibody (If more than one was used, describe in
“additional notes”) :
Concentration or dilution: 2000-250
Diluent buffer : 5%BSA or Milk Powder
Incubation time: over night
Incubation temperature: 4oC
7) Secondary antibody:
Species: Rabbit
Reacts against: Mouse
Concentration or dilution: 3000-1000
Diluent buffer: 5% BSA or Milk Powder
Incubation time: 1-2 hrs
Incubation temperature: Room Temperature
Fluorochrome or enzyme conjugate: HRP
8) Washing after primary and secondary antibodies:
Buffer: TBS-tween
Number of washes: after primary (3x5) and after Secondary (3x15)
9) Detection method: By using ECL reagent.
10) How many times have you run this staining? More than 4 times
Do you obtain the same results every time? YES
What steps have you altered to try and optimize the use of this antibody?
By Optimizing blocking steps by BSA and milk powder, amount of samples
from 20-50ug, Primary Antibody dilutions from 2000-250, Secondary
antibody dilution from 3000-1000
Document attachment: Attaching images of your blot is strongly
recommended and can greatly speed up our investigation of your problem.
 Nothing on the images…..
Regards,

Read More
Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from ab54532. The details you have kindly provided will provide us with vital information for our monitoring of 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. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial.
Pleas note that ab54532 is not tested for IP and we do not know if it is suitable.
I apologise for the inconvenience and am pleased to offer you a free of charge replacement (also possible with an alternative antibody; unfortunately, we do not have an anti-Cofilin antibody that is tested for IP) or credit note in compensation.
Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Question
Answer

Merci pour votre appel et votre patience.
Le laboratoire vient de me confirmer que ils n'ont pas fait un "epitope mapping" sur cet anticorps, et donc que l'épitope de cet anticorps n'est pas connue. Effectivement, l'immunogène était toute la protéine Cofilin avec un tag, et l'épitope peux donc être théoriquement n'importe ou sur la protéine.
Je suis désolée de ne pas pouvoir vous aider plus en cette occasion. N'hésitez pas à nous recontacter si vous avez des autres questions.

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Answer

Thank you for contacting us. I am sorry that this new vial of 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

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 for one vial of ab75510. 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.  

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Answer

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 for one vial of ab75510. This product is currently out of stock. You should expect to receive your shipment on or around Monday December 12. 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 your enquiry. I am sorry this vial of ab54532 is giving you trouble. Here is a link to another anti-Cofilin antibody you may be interested in: https://www.abcam.com/Cofilin-antibody-ab29038.html As a reminder, please check the datasheet for ab29038 to be certain it will suit your needs. We do have other anti-Cofilin antibodies if this product will not be suitable. I hope this helps. When you get a chance please let me know which antibody you would like as a replacement and what purchase order number or Abcam order reference number was associated with the purchase of ab54532 so I can process your request.  

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1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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