Recombinant Anti-Myosin Light Chain 2 antibody [EPR3741] (ab92721)

Thank you for contacting us with your questions.

The IHC images on the datasheets for ab92721 and ab137063 were produced in the Epitomics lab, and they weren't taken from publications so there are no references. Ab92721 was used in one publication-

http://www.ncbi.nlm.nih.gov/pubmed/22037191?dopt=Abstract

We only have full size vials of our antibodies, so we don't have samples to send out. However, we do have a promotion on rabbit monoclonal antibodies that is valid until December 16. For this promotion, if you purchase any primary antibody you can receive a free rabbit monoclonal. Since both of these antibodies are rabbit monoclonals (also called RabMabs), you could purchase one and then receive the other for free. If you'd like to take advantage of this offer, please include the code RABMAB-XBSMG with your order.

I hope that this information will be useful, but please let me know if you have further questions or need anything else and I'll be happy to help.

Are you interested in an antibody against a different cardiac marker such as GATA4? The antibody ab86371 looks good.

Click here (or use the following: https://www.abcam.com/GATA4-antibody-6H10-ab86371.html).

The antibody ab8295, which you used before successfully, is a cardiac troponin T antibody. Do you want another vial of that, to replace the other troponin T antibody, ab33589? If not, then I suggest ab89594.

Click here (or use the following: https://www.abcam.com/Myosin-Light-Chain-2-antibody-AT3B2-ab89594.html).

I look forward to your reply.

Thank you for bringing this to our attention. If I cannot offer a useful suggestion to improve your results, and if the antibodies are covered by our guarantee, I will send replacements.

I will first need a few details of your experiment and your order.

1. Can you please tell me when you ordered the antibodies? If you have a purchase order number or the name of the person who placed the order, that will help me find it in our records.

2. Please describe your samples: what is the species and cell type or tissue, and how were the lysates prepared? Did you use protease inhibitors?

3. How much protein did you load per lane of the gel?

4. Are you confident that the transfer from gel to blot was successful?

5. What is the blocking solution? (For example, 5% milk in TBS or 1% BSA, etc.).

6. You mentioned using the antibodies as advised on the datasheets. What dilutions did you try for each antibody?

7. Are confident the secondary antibody is effective?

I look forward to your reply.

I have issued a free of charge replacement for one vial of ab92721. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

Thank you for contacting Abcam regarding ab92721 and ab89594. I have received the following protocol information from the laboratory: Western Blotting Protocol Solutions and Reagents 2X Laemmli Sample Buffer: 62.5 mM Tris-HCl (pH 6.8), 25% glycerol, 2% SDS, 0.01% Bromophenol Blue, 710 mM beta-mercaptoethanol TBST (50 mM Tris, pH 7.6, 150 mM NaCl, 0.1% Tween-20) TBST is Tris Base Saline buffer with 0.1% Tween-20 5% bovine serum albumin (BSA): Use 25 g in 500 ml TBST Goat anti-rabbit HRP antibody - suggest ab6721 Goat anti-mouse-HRP - suggest ab6789 Chemiluminesence reagents; such as ECL materials and film or imaging system for detection Protocol Run SDS-PAGE using 10-60ug heart lysate and transfer to nitrocellulose membrane. Block nitrocellulose for 1 hour at room temperature or overnight at 4°C using 5% BSA Incubate nitrocellulose with appropriate dilutions of primary antibody in 5% BSA overnight at 4°C or for 2 hours at room temperature Wash nitrocellulose with three 5-min washes using TBST. Incubate nitrocellulose with goat anti-rabbit (or goat anti-mouse) HRP antibody, diluted to 1:500 to 100,000 in 5% BSA for 1-2 hours at room temperature. Wash nitrocellulose in 3 washes of TBST, then rinse in TBS prior to addition of chemiluminesence reagents. Remove excess chemiluminescence reagent and sandwich nitrocellulose blot in any type of transparent plastic wrap (plastic cling wrap, transparent sheet protector, etc.). Acquire image using darkroom development techniques. I hope this information is helpful.  Please to not hesitate to contact us if you have any additional questions.