Anti-MMP1 antibody [3B6] (ab2461)

Hi! Does this antibody target the active collagenase only, or does it also target the proenzyme?

Regards

ANSWER:

Thank you for you enquiry.

Further to correspondence with the source of this antibody I can tell you that this antibody targets both the active form and the proenzyme. I have updated the antibody datasheet as a result.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

What bands are seen in Western blot with these antibodies?

ANSWER:

Thank you for your enquiry.

The active enzyme has a Mw of 43 KDa. Thus in the blot you wil find both the 54 KDa band of the proenzyme, and also the 43 KDa band.

I hope this information helps. Please do not hesitate to contact us if you need anything further.

I have ordered the following products:

MMP1: ab2461 MMP9: ab2463 GM-CSF: ab649 PDGFBB: ab15499 Fibronectin: ab6328

I have an important query concerning the laboratory work: Can you tell me the concentration of the protein from MMP1 and the concentration of the protein from MMP9. I need those concentrations of the IgG fraction or the concentration of the total protein as a control.

Do you have any protocols or helps ( procedure, pH, treatment,...) for the 5 antibodies?

As I am a medical student it would be wonderful if you could give me some advice.

Thank you very much in advance.

ANSWER:

You do not state in which application you plan to use the antibodies. However, you mention procedure, pH, treatment, etc. and the fact that you are a medical student, so I assume that you are going to working with IHC on paraffin embedded sections.

Where we can, we recommend a dilution to use these antibodies in each of the applications in which they have been tested - e.g. ab6328 (Fibronectin)is recommended for use in IHC-P at a 1:200 dilution. For ab15499 (PDGFBB) we recommend a 1/100 diltuion for 10 min at RT. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.

You specifically ask about ab2461 (MMP1) and ab2463 (MMP9)which are provided as tissue culture supernatant. While there is information about a +ve control and antigen retrieval on the datasheets, we do not state a recommended starting dilution on the datasheets. The IgG in tissue culture supernatant will be less concentrated than in purified products, so I'd recommend that you start with a 1:50 dilution using 100µl of working solution per slide.

I highly recommend that you take a look at the IHC protocols section of our website. Go to the homepage and in the yellow bar at the top you will see a link to "resources". At the bottom of the drop-down list is a link to protocols where you will find information on IHC - Paraffin sections, IHC - Frozen sections / Cytological preparations and antigen retrieval.

If you have any further questions once you have started to use the antibodies, please do not hesitate to contact us again.

No details of starting dilution factor for antibody on data sheet or in the 2 references given on data sheet. Could you please let me know what your starting dilution for western blotting is with this antibody - ab2461.

ANSWER:

Thank you for your enquiry. I would recommend using a dilution of 1:1000 and then optimizing based on your results. If you have any additional questions, please contact us again.