Carbonic Anhydrase IX Antibody 0.1ml

1. We are interested in Anti-Carbonic Anhydrase IX antibody (NB 100-417). We need to know how many units we need to purchase in order to stain 150 formalin fixed, paraffin embedded kidney biopsies. It depends on the amount used per slide, so could you please confirm how much antibody do you use per slide in IHC-P? The antibody?s size is 100ul, so I would like to know the amount used to calculate the units the customer would need to buy. Also, is the ab compatible with ENVISION FLEX+, HIGH pH, FOR AS?
2. Could you please let me know the immunogen peptide location for this Ab?
3. I would like to know whether the immunogen of Carbonic Anhydrase IX Antibody (NB100-417) is the cytosolic or the extracellular region??
4. I'm going to use your CA-9 Antibody for paraffin-Immunohistochemistry. Do I have to do any special pretreatments?
5. Do you have any recommendation for a blocking buffer for use of a primary antibody in Western Blot?
6. Could you please provide information on how to find positive controls for your primary antibodies?

• We are interested in Anti-Carbonic Anhydrase IX antibody (NB 100-417). We need to know how many units we need to purchase in order to stain 150 formalin fixed, paraffin embedded kidney biopsies. It depends on the amount used per slide, so could you please confirm how much antibody do you use per slide in IHC-P? The antibody?s size is 100ul, so I would like to know the amount used to calculate the units the customer would need to buy. Also, is the ab compatible with ENVISION FLEX+, HIGH pH, FOR AS?
  Unfortunately there is no number that I can provide to you as it is impossible to know for sure how many slides you will be able to stain with a vial. This will depend on your experimental conditions, your samples and how highly expressed the protein is, and what dilution you end up choosing. This antibody is recommended to be used at a 1:1000 dilution for IHC staining as this has worked well for our lab. You should order a single vial and test to determine what amount will be best for your assay rather than order too much or too little without first working with the antibody. As for your second question that is not something that our lab has tested with so I am unsure if it is compatible or not. It has a fairly normal range pH in the buffer (7-8) so most likely the functions best around that range and not at a high pH.

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• Could you please let me know the immunogen peptide location for this Ab?
  The immunogen used to create this antibody was made to a cytoplasmic area of the protein. I hope this is helpful.

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• I would like to know whether the immunogen of Carbonic Anhydrase IX Antibody (NB100-417) is the cytosolic or the extracellular region??
  The lab has informed me that the immunogen used to create this antibody was made to a cytoplasmic area of the protein.

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• I'm going to use your CA-9 Antibody for paraffin-Immunohistochemistry. Do I have to do any special pretreatments?
  Here is our protocol specific for this product: www.novusbio.com/support/protocols/protocol-specific-for-carbonic-anhydrase-ix-antibody-NB100-417.html ?However, we recommend antigen retrieval to ensure the epitope is not masked, and we also recommend that you add a permeablizing agent to access the epitope in the nucleus.

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• Do you have any recommendation for a blocking buffer for use of a primary antibody in Western Blot?
  The blocking buffer that is often employed in our Western Blot QC validation is either 3% BSA, or 5% NFDM (Non-fat Dry Milk). In certain cases a mixture of both BSA and NFDM will be employed. Ensure that BSA is used as the blocking buffer for antibodies that target phosphorylated post translational modifications. This is to ensure that the antibody does not non-specifically bind the casein found in NFDM.

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• Could you please provide information on how to find positive controls for your primary antibodies?
  The images on our datasheets will provide detailed information on potential positive control cells and/or tissues, which have been previously tested by the lab for antibody validation. We also provide a related products tab at the top of the datasheet, which will contain useful control information where available. In addition to these tools, we recommend checking tools such as www.uniprot.org or www.proteinatlas.org to determine tissues and cell lines that highly express your protein of interest.

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