Anti-Histone H1.2抗体- ChIP Grade (ab4086)

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I am sorry that these 2 antibodies did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.

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I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx for1 vial of ab4086 with a new lot. As for ab5212, order xxx has been arranged with 1x vial oflot 863574. Theestimated delivery date for this vial is May 25th. I apologize about this delay.

Please let me know how each lot is working out for you and I will send you the second vial for each antibody.

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, and look forward to hear back from you.

Thank you very much for your telephone enquiry.

I am sorry to confirm that to our knowledge, the two histone H1.2 antibodies we have are regrettably not tested and guaranteed in mouse or frozen sections (IHC-Fr).

ab17677 Anti-Histone H1.2 antibody
Rabbit polyclonal
ICC/IF, IHC-P, IP, WB
Reacts with:Human

ab4086 Anti-Histone H1.2 antibody - ChIP Grade
Rabbit polyclonal
ChIP, ICC/IF, IHC-P, WB
Reacts with: Human

I have checked the alignment of the immunogen for these antibodies with the mouse Histone H1.2 protein. This is very quite low at 63%. Unfortunately, this means I would suggest there is likely to be little crossreactivity of these antibodies with the mouse histone H1.2 protein.

Therefore, unfortunately it is not possible for use to offer any testing discount on this occasion.

If I am sorry we have no suitably tested Histone H1.2 antibodies for your requirements on this occasion. If you have any further questions, please do not hesitate to contact us.

Thank you for contacting us. We are pleased to hear that you attended our seminar on the 16th and hope you enjoyed it. I appreciate the time you have spent in the laboratory and understand your concerns. We would like to investigate this particular case further for you and offer some suggestions to help optimise the results for your patient tissue samples. Have you tried to keep the samples on ice during the sonication procedure? For such a long sonication it is recommended to cool the samples throughout the cycles, repeatedly adding fresh ice if necessary. Also, do you cross-link your samples? The crosslinking in ChIP serves to create a snapshot of the cell at the moment when you add the formaldehyde, as it kills cells immediately and crosslinks protein to DNA. This step also has influence on the sonication efficiency. For details please find our protocol attached. May I ask if this sonication step has been optimised? Ideally, DNA fragments should be approximately 300 bp in length and it is advisable to perform a time course to determine the optimal sonication conditions. So depending on the sample type a shorter procedure may be suitable as well. Although some ChIP grade antibodies may recognize the native structure of the antigen, I would not recommend proceeding to the next step without cross-linking and sonicating the samples. I hope this information is helpful and would like to wish you good luck with your research. The technical team is always at your service, should you require further expert advice. PS: I am happy to let you know that we have a webinar on ChIP coming up (presumably in January) that might be of interest to you. Details can be found in due course on our website (https://www.abcam.com/index.html?pageconfig=tradeshow).

Thank you for your enquiry. This antibody was tested on purified core histones purchased from another company. They were purified by acid extraction precipitation from log phase untreated HeLa cells. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Thank you for your e-mail. We are very pleased to hear that you are satisfied with the antibody ab4086. I enclose our recommended protocol for blocking, please do not hesitate to contact us again if you need further information. In principle, a small volume of antibody (e.g., 1-5 ul) is first reacted with excess peptide (5-50 fold over the antibody; e.g., 1 ug antibody reacted with 5-50 ug peptide; exact amounts determined by titration) to neutralize it. The neutralized antibody can no longer subsequently bind to another antigen (a band of interest) or staining pattern. So the band(s)/staining that is competed by the antigen/peptide is supposedly specific. If more than one band disappears by peptide/antigen competition then those bands have the antigenic determinants and could be considered either fragments of the large antigen or multimer. 1. Determine the amount of antibody that is needed for 1 strip (e.g., 2 ml). For example, an antibody has given desired bands at 1:1000 dilution. So you will need 1 ul/ml antibody (2-ul antibody for 2-ml antibody solution). If an antibody were used at 1:5000 dilution then you would only need 0.2 ul/ml (use 2 ul of 1:10 dilution for better accuracy). 2. Take 2-ul antibody (or as needed) in 100 ul saline/PBS. Make 2 tubes. Add antigen/peptide solution (10-50 ug peptide or antigen added in 10-100 ul). Add same volume of saline/PBS (no peptide/antigen) to the other tube labeled as -peptide or No peptide. Mix gently. 3. Incubate both tubes at 37oC for 1-2 hrs or 2-24 hrs at 4oC. 4. Centrifuge the tubes for 15 min at 4oC in a microfuge (10-15000 rpm) to pellet any immune complexes. Carefully remove the supernatant. If no visible pellet is seen than just leave approx. 5-10 ul at the bottom to avoid disturbing invisible immune complexes. If you do not centrifuge the solution, it may give high background. 5. After centrifugation, make up the volume of supt. to 2 ml (or what is necessary depending upon the initial antibody taken) with buffer (PBS-Tween with or without BSA or milk or any buffer that was used initially). Use both antibodies (with and without antigen/peptide) for Western blotting or immunolocalization. 4. Observe the bands/staining that disappears. We have tested ab4086 specificity by mixing it at 1/500 with 1µg/ml of peptide Histone H1.2 ab16936.