Dear Kate,
Thank you for your help with ab6992 anti-human CTGF antibody.
I did the first ab6992 western blot trial using REH (positive for CTGF mRNA expression), and Raji (negative for CTGF mRNA expression) and I had a band in both of them as you can see in the attached image.
Do you know if the antibody has cross-reactivity to other proteins such as Nov or CYR61?
Thank you for your time and support.
Regards,
Abdul
Abdul Alsaleh
mailto:abdul.alsaleh@otago.ac.nz
From: Gwenn Le Mee [gwenn.lemee@otago.ac.nz]
Sent: Monday, April 02, 2012 8:52 AM
To: Abdul Alsaleh
Subject: Fwd: Reply from Abcam to your enquiry regarding ab5097, ab6992, ab74671 [CCE3646216]
Begin forwarded message:
From: <mailto:technical@abcam.com>
Date: 31 March 2012 2:12:13 AM NZDT
To: <mailto:gwenn.lemee@otago.ac.nz>
Subject: Reply from Abcam to your enquiry regarding ab5097, ab6992, ab74671 [CCE3646216]
http://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header
Dear Gwenn
Thank you for your reply.
I can confirm the following information for these two antibodies:
ab5097Anti-CTGF antibody
This antibody is testedin WB and IHC on paraffin embedded sections (IHC-P). All testedandguaranteedapplications are listed on the product datasheets and this information is updated as soon as we receive new information. I am sorry this is not tested inin flow cytometry or IHC-Fr (frozen sections).
Any publications that we are aware of will be listed on the datasheet. We have a dedicated team who spend time reviewing the literature to find articles that have used our products. However, in many cases, regrettably we do not have access to all the articles mentioning our products, or subscriptions to all the journals that publish them. I'm sorry there are no further publications we are aware offor ab5097.
ab6992 Anti-CTGF antibody
I can confirm thatall the information we have is on the datasheet and we regrettably have no data fromhaemopoietic cells for this antibody. The antibody is tested and guaranteed for WB, IHC-P, IHC-Fr, but not for flow cytometry.
Regarding the WB results shown on the datasheets forab51704 and ab59681 are from recombinant. or fusion protein samples, which is why the molecular weight is not as expected size for CTGF.
If you would like to test ab6992 or ab5097 in flow cytometry, please contact me again 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 ashttp://Amazon.comgift certificates.
To find out more about our Abreview system, please see the following link:
http://www.abcam.com/abreviews
I hope this information is helpful. Please do not hesitate to contact me for any further advice or information.
Help us improve our service.
http://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3646216
Best regards,
Kate
Kate Hayes
Scientific Support Supervisor
Abcam plc
http://www.abcam.com
Your original inquiry to Abcam:
Dear Kate,
Thank you for your answer.
I am interested in using one of the first 2 antibodies (ab5097 and ab6992) as there is less informations on the datasheet for the other 2, and both show a band at 13kDa rather than 38kDa.
As ab5097 was made against human CTGF we would naturally go for that one, but
The literature search showed only two articles using ab5097 antibody, both tested the antibody on human tissue (Van Der Laar et al., tested the antibody on human endothelial cells from the aortic wall, and Bellahence et al., tested the antibody on breast cancer cell lines [MDA-MB-31, B02]). Are you aware of any other study that was done on haemopoietic cells?
Ref: Van Der Laar IMBH et al. Mutations in SMAD3 cause a syndromic form of aortic aneurysms and dissections with early-onset osteoarthritis. Nature Genetics 2011: 43;121-126.
Bellahcene etal. Transcriptome analysis reveals an osteoblast-like phenotype for human osteotropic breast cancer cells. Breast Cancer Research 2007: 101(2);135-148.
On the other hand, ab6992 antibody was used by many studies, and some of them showed good and clear results on western blot and IHC. However, most of them are on rat/mice and we couldn’t find any study that used this antibody on haemopoietic cells, are you aware of any?
We are planning to use one of them on the following applications:
1.Western Blotting.
2.Immunohistochemistry.
3.Fluorescent Activated Cell Sorting (FACS).
It would be great if you could advice me on which one would be the most appropriate for my study.
Finally, what is the price of both antibody?
Thank you for your time and support.
Regards,
AbdulAlsaleh
On 29/03/2012, at 1:45 AM, <mailto:technical@abcam.com>
<mailto:technical@abcam.com> wrote:
http://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header
Dear Gwenn
Thank you for your enquiry.
I can confirm we have the following CTGF antibodies tested in WB and either IHC-Fr or IHC-P.
ab5097 CTGF antibody
Rabbit polyclonal
Tested in: ICC/IF,IHC-P, WB
Reacts with: Human, Rat
Clickhttp://www.abcam.com/index.html?datasheet=5097(or use the following:http://www.abcam.com/index.html?datasheet=5097).
ab6992 CTGF antibody
Rabbit polyclonal
Tested in: ICC/IF, IHC-FoFr, IHC-Fr, IHC-P, IP, WB
Reacts with: Human, Mouse, Rat, Sheep
http://www.abcam.com/index.html?datasheet=6992(or use the following:http://www.abcam.com/index.html?datasheet=6992).
ab51704 CTGF antibody
Mouse polyclonal
Tested in: ELISA, IHC-Fr, IHC-P, WB
Reacts with: Human, Mouse
Clickhttp://www.abcam.com/index.html?datasheet=51704(or use the following:http://www.abcam.com/index.html?datasheet=51704).
ab59681 CTGF antibody - Carboxyterminal end
Rabbit polyclonal IHC-Fr, IHC-P, WB
Reacts with: Human, Mouse, Rat
http://www.abcam.com/index.html?datasheet=59681(or use the following:http://www.abcam.com/index.html?datasheet=59681).
I would like to reassure you that our antibodies have all been tested successfully and would be covered by our guarantee in the applications listed on the datasheets. Therefore, any CTGFantibody tested in the species and application you are using should be suitable for yourapplications. Please check if you are using IHC on frozen (IHC-Fr) or paraffin (IHC-P) sections and I would recommend to select an antibody tested in that application.
Regrettably, I am sorry we have not CTGF antibodies tested and guaranteed in flow cytometry. All tested applications and species are listed on the datasheet, and these are updated as soon as we receive new information. If you would like to test one of these antibodies in flow cytometry, please let me know before you purchase as you may be eligible for our testing discount program.
With regards towhich antibody to recommend,although we produce many in-house antibodies, we obtain other antibodies from a wide range of sources. Therefore, comparison experiments will not often have been done. If you wish to be more certain, I can suggest choosing the antibody that has been best characterized. By this I mean the antibody that has been tested in the most applications and has the most data available (on the datasheet).
I hope this will be helpful. Should you have any further questions please do not hesitate to contact me.
Help us improve our service.
http://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3638002
Best regards,
Kate
Kate Hayes
Scientific Support Supervisor
Abcam plc
http://www.abcam.com
Your original inquiry to Abcam:
Hi,
I am a master student at Otago University (Pathology Department/Dunedin) in New Zealand. I am looking for primary anti-CTGF antibodies that bind to human CTGF (Connective Tissue Growth Factor) protein.
I need to use it for the applications below:
1.Western Blotting.
2.Immunohistochemistry.
3.Possibly Fluorescent Activated Cell Sorting (FACS).
I will be identifying CTGF protein from human haemopoietic cells (fetal lymphoblasts, and fetal lymphocytes).
I noticed a number of anti-CTGF antibodies on your website, and I am wondering if you could recommend me one of them.
Thank you for your time and assistance.
Regards,
Abdul Alsaleh
Abcam Customer Services and Scientific Support Team
http://www.abcam.com/index.html?pageconfig=technical&utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Body
[CCE3638002]
Discover moreathttp://abcam.com
------------
Dr Gwenn LE MÉE
Assistant Research Fellow
Department of Pathology
Dunedin School of Medicine PO-Box 913
University of Otago
Dunedin 9054
New Zealand
mailto:gwenn.lemee@otago.ac.nz
0064 (0)3 470 3455
Abcam Customer Services and Scientific Support Team
http://www.abcam.com/index.html?pageconfig=technical&utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Body
[CCE3646216]
Discover moreathttp://abcam.com
------------
Dr Gwenn LE MÉE
Assistant Research Fellow
Department of Pathology
Dunedin School of Medicine PO-Box 913
University of Otago
Dunedin 9054
New Zealand
mailto:gwenn.lemee@otago.ac.nz
0064 (0)3 470 3455 |
ANSWER: |
|
Thank you for your reply. I am sorry to hear you have some concerns regarding the results from this antibody.
Reviewing the image, there is a band at the correct size. This is quite faint, but with optimization it is probable that this could be improved. For example:
1. Increasing the incubation time to overnight 4oC. This can often provide more specific and efficient staining if this has not already been tried.
2. Try increasing the concentration of antibody,
3. Checking the transfer of protein to the membrane and quality of the sample using a loading control.
With regards to the results from the Raji cells, the following publication indicates there is CTGF expression in 25% of Raji cells. Could you confirmhow thesecells were selected as a negative control? Are these cells siRNA knockouts? Although the RT PCR indicates no CTGF mRNA expression, this does not give an accurate indication of the possible presence of residual protein.
http://ajcp.ascpjournals.org/content/133/2/271.full
Connective Tissue Growth Factor Is Expressed in Malignant Cells of Hodgkin Lymphoma but Not in Other Mature B-Cell Lymphomas
Anna Birgersdotter, et al
(2010) American Journal of Clinical Pathology, 133, 271-280
I can recommend a further literature search to obtain more information.
With regards to your concerns about cross reactivity, I am sorry this antibody has not specifically been tested for cross reactivity to other proteins. However, I can confirm that the mouse immunogen sequence has 47% alignment to human CRY61 protein and 15% alignment to human Nov protein. These alignments are very low, and it is therefore very unlikely that the antibody will cross react with these other targets. Is there anyreason you are concerned about the concerned about these proteins in particular?
I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forWB and human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.
Therefore, if you are still concerned about the results, I would like to investigate this further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.
Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.
Order Details
Antibody code:
Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background
Lot number
Purchase order number
or preferably Abcam order number:
General Information
Antibody storage conditions (temperature/reconstitution etc)
Description of the problem (high background, wrong band size, more bands, no band etc.)
Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Amount of protein loaded
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Detection method (ECL, ECLPlus etc.)
Positive and negative controls used (please specify)
Optimization attempts (problem solving)
How many times have you tried the Western?
Have you run a "No Primary" control?
Yes No
Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?
What steps have you altered?
Additional Notes:
Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results. |
Thank you for your answer.
I am interested in using one of the first 2 antibodies (ab5097 and ab6992) as there is less informations on the datasheet for the other 2, and both show a band at 13kDa rather than 38kDa.
As ab5097 was made against human CTGF we would naturally go for that one, but
The literature search showed only two articles using ab5097 antibody, both tested the antibody on human tissue (Van Der Laar et al., tested the antibody on human endothelial cells from the aortic wall, and Bellahence et al., tested the antibody on breast cancer cell lines [MDA-MB-31, B02]). Are you aware of any other study that was done on haemopoietic cells?
Ref: Van Der Laar IMBH et al. Mutations in SMAD3 cause a syndromic form of aortic aneurysms and dissections with early-onset osteoarthritis. Nature Genetics 2011: 43;121-126.
Bellahcene etal. Transcriptome analysis reveals an osteoblast-like phenotype for human osteotropic breast cancer cells. Breast Cancer Research 2007: 101(2);135-148.
On the other hand, ab6992 antibody was used by many studies, and some of them showed good and clear results on western blot and IHC. However, most of them are on rat/mice and we couldn’t find any study that used this antibody on haemopoietic cells, are you aware of any?
We are planning to use one of them on the following applications:
1. Western Blotting.
2. Immunohistochemistry.
3. Fluorescent Activated Cell Sorting (FACS).
It would be great if you could advice me on which one would be the most appropriate for my study.
Finally, what is the price of both antibody?
Thank you for your time and support.
Regards, |
ANSWER: |
|
Thank you for your reply.
I can confirm the following information for these two antibodies:
ab5097 Anti-CTGF antibody
This antibody is tested in WB and IHC on paraffin embedded sections (IHC-P). All tested and guaranteed applications are listed on the product datasheets and this information is updated as soon as we receive new information. I am sorry this is not tested inin flow cytometry or IHC-Fr (frozen sections).
Any publications that we are aware of will be listed on the datasheet. We have a dedicated team who spend time reviewing the literature to find articles that have used our products. However, in many cases, regrettably we do not have access to all the articles mentioning our products, or subscriptions to all the journals that publish them. I'm sorry there are no further publications we are aware of for ab5097.
ab6992 Anti-CTGF antibody
I can confirm thatall the information we have is on the datasheet and we regrettably have no data fromhaemopoietic cells for this antibody. The antibody is tested and guaranteed for WB, IHC-P, IHC-Fr, but not for flow cytometry.
Regarding the WB results shown on the datasheets forab51704 and ab59681 are from recombinant. or fusion protein samples, which is why the molecular weight is not as expected size for CTGF.
If you would like to test ab6992 or ab5097 in flow cytometry, please contact me again 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:
http://www.abcam.com/abreviews
I hope this information is helpful. Please do not hesitate to contact me for any further advice or information. |
