Anti-Osteopontin antibody (ab8448)

our customer prefers to get a new vial of ab8448 (different lot).

Please confirm when are you planning to ship it.

Kind regards

ANSWER:

Thank you for kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

As requested, we are pleased to arrange a free of charge replacement. I can confirm that 1 vial of ab66587 from a different lot numberhas been added to your order number ####. This has Abcam order reference ######.

I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

please find here our customer's reply:

1. Please confirm what size are the bands? I'd appreciate if you are able to provide an image, including molecular weight markers, which would help me to assess the results.

2. Has the quality of the sample been assessed using a loading control?

I have not a loading control.

3. Has the sample been reduced and denatured?

Yes, sample has been reduced and denatured

4. Is the current vial of secondary antibody working well with other primary antibodies? I can recommend it would be beneficial to run a no primary control to assess if the secondary is binding non specifically. The concentration of the secondary may require optimization.

Secondary antibody works well with other primary antibodies, and I also tried to dilute secondary antibody but results have been similar.

I hope this information help you.

Kind regards

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 be pleased to arrange a free of charge replacement or credit note in compensation.

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

Product code: 8448
Lot number: GR52573-4
1)Antibody storage conditions: Reconstitued antiboby has been store at -20oC.
2)Description of the problem: More bands
3)Sample: Protein have been extrated from mice dermal fibroblast and it has been done a total lysis
4)Sample preparation: Cells were washed several times with phosphate-buffered saline and homogenized in RIPA buffer in presence of protease inhibitors. Cellular lysates were centrifuged at 15,000 rpm for 20 min to clear cell debris, and supernatants were collected and stored at -80C until analysis.
5)Amount of protein loaded: 30 ug proteins
6)Electrophoresis/Gel conditions: 10% polyacrylamide gel, under reducing conditions
7)Transfer and blocking conditions:Transferred to nitrocellulose. Membrane was blocked in TBS+0.1% Tween 20 + 5% non fat dry milk for 1 hr at room temperature
8)Primary Antibody: Antibody was diluted in TBS-0.1% Tween 20 + 2.5% non-fat dry milk at 1:1000 and incubation time was overnight at 4°C. 3 wash steeps and incubation of nitrocellulose with
9)Secondary Antibody: Secondary antibody (no Abcam antibody) was diluted in TBS-Tween 20 + 2.5% non-fat dry milk at 1:5000 for 1 hr.
10)Detection method: Western blots were visualized using Super Signal West Pico
11)Positive and negative controls used (please specify): No
12)How many times have you tried the Western?: 3 times
13)Have you run a "No Primary" control?: No
14)Do you obtain the same results every time?: Yes and the number of bandes were always the same.
15)What steps have you altered?: I modified concentration of antibody both primary (1:2500) and the incubation time (instead overnight I tried only 1hr)

ANSWER:

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab8448 is tested and covered by our 6 month guarantee for use inWB and mouse samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results. I would also appreciate if you can confirm some further details:

1. Please confirm what size are the bands? I'd appreciate if you are able to provide an image, including molecular weight markers, which would help me to assess the results.

2. Has the quality of the sample been assessedusing a loading control?

3. Has the sample been reduced and denatured?

4. Is the current vial of secondary antibody working well with other primary antibodies? I can recommend it would be beneficial to run a no primary control to assess if the secondary is binding non specifically. The concentration of the secondary may require optimization.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details and the image.

Dear technical team,
Our customer had purchase order those 4 antibodies today and he is going to IHC-P with mouse bone tissue.
Please confirm bone tissue also applicable to those antibodies.
(ab8448, ab93876, ab15580, ab108337)

ANSWER:

Thank you for contacting us.

According to the expression profile of these proteins:

Osteopontin is highly expressed in mouse bone tissue:

http://www.ncbi.nlm.nih.gov/UniGene/ESTProfileViewer.cgi?uglist=Mm.288474

Ki67 would show moderate staining in IHC with bone tissue:

http://www.proteinatlas.org/ENSG00000148773

Osteocalcin is also abundant in this tissue: http://www.proteinatlas.org/ENSG00000242252 as well as Bone Alkaline Phosphatase: http://www.ncbi.nlm.nih.gov/UniGene/ESTProfileViewer.cgi?uglist=Mm.288186

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

This customer raised an enquiry about ab17989, ab8448 and ab13418. He has ordered the ab17989, ab8448 and ab13418 recently, the Lot number as follow: ab17989--748480 ab8448--gr52573-2 ab13418--gr54833-1 Would you please offer the COC for him? Thanks for your kindly help Best regards

ANSWER:

Thank you for contacting us.

Please find attached the requested Certificate of Compliance

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