This Ki67 antibody is useful for Immunohistochemistry on frozen and paraffin-embedded sections and Immunocytochemistry/Immunofluorescence. There have been mixed results in Western blot; however, NB500-170 has been used successfully Western Blog reported by a customer review. *Formalin fixed paraffin embedded tissue sections require high temperature antigen unmasking with 10 mM citrate buffer, pH 6.0 prior to immunostaining. This antibody will not work without optimal antigen retrieval. This is probably the most critical step. NOTE: We suggest an incubation period of 30 minutes at room temperature and to use DAB to stain the protein (immunofluorescence may give problems as the protein is nuclear).
Originally discovered employing mouse monoclonal antibody against a nuclear antigen from Hodgkin's lymphoma-derived cell line, this non-histone protein was named Ki67 after researcher's location (Gerdes and colleagues), Ki for Kiel University in Germany and 67 referring to the clone number on the 96-well plate. It interacts with KIF15 as well as MKI67IP, and is suggested to be involved in cell cycle regulation. Ki67 is a large protein with expected molecular weight of about 395 kD and has a very complex localization pattern within the nucleus, one which changes during cell cycle progression. Its expression occurs specially during late G1, S, G2 and M phases of the cell cycle, while in cells undergoing G0 phase, Ki67 remains undetectable. Ki67 undergoes phosphorylation/dephosphorylation during mitosis, is susceptible to proteases and its structure implies that its expression is regulated by proteolytic pathways. Ki67 is associated with nucleolar DFC (dense fibrillary component) and its regulation appears to be tightly controlled (estimated half life is 60-90 min, regardless of the cell position in the cell cycle), presumably by precise synthesis and degradation systems involving proteasome, a protease complex. Due to its association with cell divison process, Ki-67 is routinely used as cellular proliferation marker of solid tumors as well as certain hematological malignancies, and a correlation has been demonstrated between Ki-67 index and the histopathological grade of cancers.
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FAQs for Ki-67/MKI67 (4)
Could you please let us know if you have the antibody which may have cross-reactivity to hamster tissue in IHC(paraffin or frozen)? Also, please tell us reference information using your products to hamster tissue.
Thank you for your patience. The only information I was able to gain is that the protein has 61% homology with the chinese hamster. Our lab only recommends use if the % homology is 85% or above so I would not recommend our Ki67 antibodies for your customer. However if they are willing to still test this they would become eligable for our Innovators Reward Program in which they would gain a 50% refund and a 50% discount on a future purchase. All you would need to do is email us at email@example.com. Here is the link to all of our Ki67 antibodies.
What is the IF protocol which was used to obtain the IF image on the datasheet?
Our lab used our general ICC protocol to obtain the image seen on our website.
In the immunohistochemistry paraffin section protocol....the PBS buffer....should it be at certain pH?
For IHC application, different labs cites the pH of PBS buffer in the range of 7.1 - 7.6 but most commonly used pH is pH 7.4 (this is what we use in our lab). You may use this PBS (pH 7.4) for making permeablization buffer, antibody diluent buffers and for making wash buffer. For more on the protocol that we use in our lab and for IHC-P troubleshooting suggestions, you may visit: IHC-P Protocol and IHC-P Troubleshooting.
I was wondering what does Format 7C mean as far as anitbodies go?
7C is a fluorophore - it is useful for FLOW. The details of the fluor are (A=425, E=500)