Anti-Actin antibody [AC-40] (ab11003)

Could you please advise the lot & expiry for the new lot of ab11003?

ANSWER:

The new lot of ab11003 is currently in transit from the production lab to our stock and there is no lot number allocated yet, but I can confirm that this new lot is different from the lot your customer has received originally.

I will send you the lot number as soon as it is created.

We do not have set expiration dates for our products.Aliquotted, not diluted and stored at -20ºC, antibodies, includingab11003is stable for up to 5 years. Up to 10 years when stored at -80ºC. It is very importantto avoid repeated freeze / thaw cycles.We guarantee all of our products to work for at least 120 days from the date of purchase when stored correctly.For more information on antibody storage and stability, please visit our Antibody Storage Guide :http://www.abcam.com/index.html?pageconfig=resource&rid=10795 .

Our customer would like to take back the same item, AB11003. However, they would like to have a another lot other thanGR2058. Please revert if this is acceptable.

ANSWER:

Thank you for your reply.

Unfortunately we do not have any other lot of ab11003 in stock.

Would your customer be interested in trying an alternative anti-actin from the following list (excluding ab11003 of course) : http://www.abcam.com/index.html?pageconfig=searchresults&search=Actin&pt=1&sk=conj&sv=0&sn=Unconjugated&l=2&fViewMore=1

Thank you.

Resend
1) Abcam product code ab11003

2) Abcam order reference number or product batch number

Cat no:AB11003

Lot no.: GR2058-9

3) Description of the problem: no band / high background (many non-specific bands) / wrong band size / other? Some are high background; some have no bands

4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…): brain lysate

Species : mice

Lysis buffer : RIPA

Protease inhibitors: Roche complete

Phosphatase inhibitors : Na-β-glycerophosphate, Na Orthovanadate

Reducing agent : b-mercapethanol

Boiling for ≥5 min? yes

Protein loaded ug/lane or cells/lane : 250

Positive control : c57 mouse brain lysate; Cre-transfected HEK

Negative control : HEK





5) Percentage of gel

Type of membrane: SDS-PAGE 10%

Protein transfer verified : Yes, all the ladders pass through

Blocking agent and concentration 5% BSA

Blocking time 1 hr

Blocking temperature Room Temperature





6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution 1:1000

Diluent buffer : 5% BSA

Incubation time Overnight

Incubation temperature: 4 deg C



7) Secondary antibody:

Species: Goat

Reacts against: Mouse

Concentration or dilution 1:5000

Diluent buffer 1% BSA in PBST-0.1%

Incubation time 1 hour

Incubation temperature: Room Temperature

Fluorochrome or enzyme conjugate: HRP





8) Washing after primary and secondary antibodies:

Buffer PBST-0.1%;

Number of washes 3 times



9)Detection method: 1 Super Signal West Pico Stable Peroxide Solution : 1mL Enhancer Solution or

Femto







10) How many times have you run this staining?

Do you obtain the same results every time? 7; 2 by other colleagues
What steps have you altered to try and optimize the use of this antibody? I changed lysis buffer and added in c57 mice and HEK cell for comparison. Actin antibody was the last thing I thought could be wrong until it was verified by other colleagues.

ANSWER:

Thank you for filling in the questionnaire and for reporting this problem to us. We will use the details you sent about your protocol in order to investigate the source of the problem.

I have looked through your protocol, which looks fine to me. It seems that you have received a faulty tube or lot of ab11003. I appreciate the time that has been spent on these experiments and would be pleased to arrange in compensation a credit note or areplacement with an alternative anti-actin from our catalog. The link to thelist of Abcamanti-actin antibodies is : http://www.abcam.com/index.html?pageconfig=searchresults&search=Actin&pt=1&sk=conj&sv=0&sn=Unconjugated&l=2&fViewMore=1

I look forward to hearing from you with details of how you andyour customerwould like to proceed.

One of our customer purchased the above item and had faced difficulties with working on the anti body. I have attached pictures, displaying the results of the experiment.

ANSWER:

Thank you for contacting us.

I am sorry to hear your customeris 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.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

Our end user want to use loading control. but a8226 data sheet describe that " Does not cross-react with adult cardiac, smooth, or skeletal muscle actin. " The sample of end user is myoblast, so they wonder they can use this antibody.

And another antibody ab 8224 is not describe like ab8226.

And then they wonder they can use ab11003 alpha actin.

Could you recommend best loading control for myoblast sample?

I wait your reply.

ANSWER:

Thank you for your enquiry. Ab8226 does not cross-react with ADULT cardiac, smooth, or skeletal muscle actin. Myoblasts are immature muscle cells and looking through the literature it does seem that some people use actin as a loading control. Your customer may want to try a tubulin loading control. However, I do not want to recommend a product that is unsuitable and suggest that you refer to the latest literature in this field.