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Lot # 1513515-COA

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Product FAQ

I know I need to block my samples against non-specific protein binding of my antibodies, but which blocker should I use?

Answer

For blocking cells or tissues you may use 2-5% bovine serum albumin (fraction V, defatted BSA) or 5-10% normal serum of the species matching the host species of the secondary antibody. Other options include a mixture of BSA and serum or other purified proteins. We offer a ready-made blocking solution, BlockAid Blocking Solution (Cat. No. B10710), which is not species-specific. For cell or tissue samples, avoid the use of non-fat dry milk as a blocking agent as it contains a high level of phosphoproteins, histones, and biotin.

Answer Id: E14772

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Lot # 1123327-COC

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  • 300400-1000-TS

Product FAQ

What fixative should I use to fix my cells or tissues for antibody labeling?

Answer

Aldehyde-based fixatives (e.g., formaldehyde, glutaraldehyde) crosslink various cellular components, which helps retain proteins and cell morphology, but some antigens can be masked by the crosslinking, requiring antigen retrieval methods to unmask the antigen binding sites. Also, aldehyde-based fixation requires permeabilization to allow entry of antibodies. Organic solvents such as methanol and acetone condense proteins and permeabilize the cells, but cell morphology can be compromised. Surface antigens can be labeled without fixation.

Answer Id: E14773

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Lot # 1613814

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  • 73684106

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Lot # TA2502176

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Lot # 1137013-COC

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Product FAQ

What concentration of my antibody should I use for cell analysis?

Answer

An optimal concentration may be between 1-10 μg/mL for cell and tissue labeling for microscopy, or 0.2-5 μg/mL for flow cytometry. A range of concentrations should be tested to determine what is optimal.

Answer Id: E14774

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Lot # 1139601-COA

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  • PO0291A-TS

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Lot # TB2527756

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Lot # 2065-COA

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  • MP1331-TS

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Lot # 1507019

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Lot # 1094475-COA

Catalog #
  • BO1100Z-TS

Product FAQ

I have a very low-abundance antigen. How can I amplify my signal?

Answer

A common method for amplifying antibody detection is biotin-streptavidin detection, where a biotinylated secondary antibody is combined with subsequent labeling with a dye-conjugated streptavidin. This will amplify the signal by approximately 2-8 times, but endogenous biotin must be blocked beforehand. Another option is to use tyramide-signal amplification, where a horseradish peroxidase conjugate is used with a dye-labeled tyramide. This will amplify the signal by approximately 10-20 times, but endogenous peroxidase will need to be blocked. A final option may be to use a Qdot nanoparticle antibody or streptavidin conjugate, which can yield a signal as much as 40 times higher than a standard organic dye conjugate, depending on the Qdot color.

Answer Id: E14775

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Lot # 1111831

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  • 455-0500-LSG