Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade (ab1012)

Dear All,



Sending you attached a list of the tendered items. You can see the products at page 3-5. Please give all of the items a check and let us know which items are best available with Abcam.



Early response requested.



Sincere regards,

ANSWER:

Thank you for your inquiry.

We have a number of products that are comparable with the products on the list you sent us. For future inquiriesI would like to direct you to the search facility on our website (www.abcam.com).

Please enter the name of the protein into the search box at the top of the home page. A list of products will be generated which can be further refined using the filters on the left hand side of the page. Using these filters will help you to identify products that have been tested in the applications and species you are interested in.

Please find a list of the product code and the links to the respective datasheets below:

ab1012 (Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade)

http://www.abcam.com/index.html?datasheet=1012 (or use the following: http://www.abcam.com/index.html?datasheet=1012).

ab12209 (Anti-Histone H3 (tri methyl K4) antibody [mAbcam12209] - ChIP Grade)

http://www.abcam.com/index.html?datasheet=12209 (or use the following: http://www.abcam.com/index.html?datasheet=12209).

ab6002 (Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade)

http://www.abcam.com/index.html?datasheet=6002 (or use the following: http://www.abcam.com/index.html?datasheet=6002).

ab124462 (Anti-Flag Tag antibody)

http://www.abcam.com/index.html?datasheet=124462 (or use the following: http://www.abcam.com/index.html?datasheet=124462).

ab6939 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Cy3 ®), pre-adsorbed)

http://www.abcam.com/index.html?datasheet=6939 (or use the following: http://www.abcam.com/index.html?datasheet=6939).

ab96899 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed)

http://www.abcam.com/index.html?datasheet=96899 (or use the following: http://www.abcam.com/index.html?datasheet=96899).

ab96919 (Donkey anti-Rabbit IgG H&L (DyLight® 488) secondary antibody)

http://www.abcam.com/index.html?datasheet=96919 (or use the following: http://www.abcam.com/index.html?datasheet=96919).

ab96879 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (DyLight® 488), pre-adsorbed)

http://www.abcam.com/index.html?datasheet=96879 (or use the following: http://www.abcam.com/index.html?datasheet=96879).

ab96871 (Goat anti-Mouse IgG H&L (DyLight® 488) secondary antibody)

http://www.abcam.com/index.html?datasheet=96871 (or use the following: http://www.abcam.com/index.html?datasheet=96871).

ab97265 (Goat anti-Mouse IgG Fc (HRP) secondary antibody)

http://www.abcam.com/index.html?datasheet=97265 (or use the following: http://www.abcam.com/index.html?datasheet=97265).

ab7481 (Goat Serum)

http://www.abcam.com/index.html?datasheet=7481 (or use the following: http://www.abcam.com/index.html?datasheet=7481).

ab7487 (Rabbit Serum (sterile))

http://www.abcam.com/index.html?datasheet=7487 (or use the following: http://www.abcam.com/index.html?datasheet=7487).

ab46666 (Herring sperm ssDNA)

http://www.abcam.com/index.html?datasheet=46666 (or use the following: http://www.abcam.com/index.html?datasheet=46666).

ab52579 (Ribonuclease A protein (Active))

http://www.abcam.com/index.html?datasheet=52579 (or use the following: http://www.abcam.com/index.html?datasheet=52579).

ab73430 (DNase I protein (Active))

http://www.abcam.com/index.html?datasheet=73430 (or use the following: http://www.abcam.com/index.html?datasheet=73430).

ab64220 (Proteinase K)

http://www.abcam.com/index.html?datasheet=64220 (or use the following: http://www.abcam.com/index.html?datasheet=64220).

ab65621 (Protease Inhibitor Cocktail)

http://www.abcam.com/index.html?datasheet=65621 (or use the following: http://www.abcam.com/index.html?datasheet=65621).

ab116028 (Prism Ultra Protein Ladder (10-245 kDa))

http://www.abcam.com/index.html?datasheet=116028 (or use the following: http://www.abcam.com/index.html?datasheet=116028).


I hope this information is helpful to you. If you have specific questions about the products identified during the search, please do not hesitate to contact me.

Thanks for your help with that order. I have another question for an additional project we are initiating requiring lots of different antibodies. This is a clinical diagnostics projects, and all antibodies we will use for that need to be CE certified for diagnostic purposes. Since I did not find any information on this on your website, I wanted to ask you if you have any CE certified antibodies directed against these (human) proteins:

ANSWER:

Thank you for contacting Abcam.

All of our products are for research purposes only and are not certified for clinical research applications.

I am sorry that I could not be more helpful in this matter.

Hi Jeremy,   Thank you so much for your kind suggestions. Today I am going to implement your suggestions. I will let you know tomorrow.  

ANSWER:

I am happy to help. Best of luck with your experiment!

Thanks for your reply.   I used the antibody concentration 1 microgram/ml in 5% BSA and I loaded the protein 5 micrograms in each lane. Could you please, suggest me how can I solve my problem.  

ANSWER:

Sorry for the late reply. I have a couple suggestions that may help improve your western blot results.

1. 5ug is quite a lot of purified protein to load on a western blot. This may result in non-specific detection of the loaded protein. I think the real specificity of this anibody may be demonstrated by loading less purified protein. I suggest a small dilution series starting at a lower mass of protein loaded. For example, load 1) 400ng 2) 100ng 3) 25ng of protein for each on the same gel.

2. Also ensure you are using BSA to block the membrane as milk has been shown to not work as a blocking reagent with ab1012.

I look forward to hearing if these suggestions improve your results.

Dear Sir/Madam,   This is with reference to one of your products [Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] (ab1012)].   The attched immunoblot demonstrates your antibody (ab1012) is false positive with unmethylated recombinant Histone H3, human (company name BPS & catalog# 52027). I had expected only core Histones, Hela from Millipore (catalog# 13-183) to be postive while recombinant Histone H3, Human (company name BPS & catalog# 52027) should be negative.   At your earliest convenience please, try to help us to resolve the problem.    FYI: The lot number of Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] (ab1012) is 940499.   Thank you very much with an anticipation.   With best, Koustav Sarkar Room# 9137, Vyas Lab Children's Hospital of Pittsburgh of UPMC

ANSWER:

Thank you for your enquiry. It does appear that the vial of ab1012 you received is reacting with unmethylated histone from BPS. This conflicts with our internal testing which shows that antibody to be largely specific for only histone H3 (tri-mehtyl K4). The only cross reactivity in our tests was slight reactivity with histone H3 (di-methyl K4).   I was just curious what dilution factor was used to dilute ab1012 for this blot and how much recombinant protein was loaded per lane.