Products:Neuroscience >> Neurotransmission >> Calcium Signaling >> Calmodulin / CaMK
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Hi Technical,
We have a customer who is interested in the above antibody and would like to know what the concentration is of the current lot you have in stock.
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ANSWER: |
Thank you for contacting us.
I have been informed by the source of the CaMKII (phospho T286) antibody ab32678 that the activity, rather than the concentration, is used to determine the package size, but the concentration of the antibody which gives the appropriate activity is between 150-250 ug/ml and this should be consistent between lots of the antibody.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. |
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I was just wondering if this antibody can detect the phoshorylated form of camkii gamma isoform as well ..... looking forward to your response... |
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ANSWER: |
Thank you for your enquiry. ab32678 was generated against the alpha/beta subunits of the protein. Further to correspondence with the source of this antibody I can tell you that its reactivity for the gamma subunit has not been tested. However, the sequence is fairly well conserved in the gamma subunit and may detect this isoform in addition. I am sorry that I cannot be of more assistance on this occasion. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
All lanes : Anti-CaMKII (phospho T286) antibody (ab32678) at 1/1000 dilution
Lane 1 : Rat brain cortex lysate.
Lane 2 : Rat brain cortex lysate preincubated with lambda-phosphatase.
Lysates/proteins at 50 µg per lane.
Predicted band size : 50 kDa
Observed band size : 50,60 kDa (why is the actual band size different from the predicted?)
Predicted molecular weight: ~50 kDa for the alpha subunit and ~60 kDa for the beta subunit of CaMKII.
These images show the phosphospecificity of this antibody.
ab32678 (2µg/ml) staining CaMKII (phospho T286) in human Brain:cortex:frontal-lateral using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of nuclear/cytoplasmic compartments within the stellate cells and the myelinated fibres of white matter region .
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
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