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BioLegend Fixation

Fixation is a process that helps to lock proteins in place on cells you plan to analyze. There can be a variety of reasons to fix your cells, including:

  • Timing/Covenience (experiments that cannot be completed in one day)
  • Safety (biohazardous samples that must be fixed/killed prior to analysis)
  • Intracellular analysis of targets (also includes permeabilization)

Fixation is a common practice, but it creates some difficulties if samples are fixed prior to staining for surface antibodies. Fixation can alter the epitope, making it difficult for your antibody to recognize its target. The information provided below is based on our in-house testing of our clones on unfixed cells (no fixation) and cells fixed with 4% paraformaldehyde prior to staining (post-fixation). We have provided data where possible. Black histograms/dot plots represent unstained samples. Purple histograms/dot plots represent staining with the indicated test antibody.

Please note that this data is based on several variables, including the types of cells analyzed, the antibody's fluorophore, and relative abundance of the antigen. Costains are also recommended to help you identify your population of interest. If you are planning to use a live/dead stain (such as Zombie Dyes), stain with them prior to fixation.

While these results will not necessarily guarantee the success or failure of staining a fixed epitope, it will at least provide you with an idea of how your samples might look.

 

Human Mouse View Tips
Specificity Clone Compatability Sample Costain Data Products
CCR8L263G8NoLysed whole blood, lymphocytesView DataView Products
CCR106588-5NoLysed whole blood, lymphocytesView DataView Products
CD3UCHT1YesLysed whole blood, lymphocytesView DataView Products
CD3OKT3YesLysed whole bloodView Products
CD3HIT3aYesLysed whole bloodView Products
CD3SK7YesLysed whole bloodView Products
CD4OKT4YesLysed whole bloodView Products
CD4RPA-T4YesLysed whole bloodView Products
CD5UCHT2YesLysed whole bloodView Products
CD6BL-CD6YesLysed whole blood, lymphocytesView DataView Products
CD7CD7-6B7YesLysed whole blood, lymphocytesView DataView Products
CD8RPA-T8YesLysed whole bloodView Products
CD8HIT8aYesLysed whole bloodView Products
CD11aHI111YesLysed whole blood, lymphocytesView DataView Products
CD11bICRF44YesLysed whole bloodView Products
CD11c3.9NoLysed whole blood, monocytesView DataView Products
CD13WM15YesLysed whole bloodView Products
CD14M5E2YesLysed whole blood, monocytesView DataView Products
CD14HCD14YesLysed whole bloodView Products
CD15W6D3YesLysed whole bloodView Products
CD15HI98YesLysed whole blood, granulocytesView DataView Products
CD163G8YesLysed whole bloodView Products
CD18TS1/18YesLysed whole bloodView Products
CD19HIB19YesLysed whole blood, lymphocytesView DataView Products
CD23EBVCS-5YesLysed whole bloodView Products
CD24ML5NoLysed whole blood, lymphocytesView DataView Products
CD25BC96NoLysed whole bloodView Products
CD27O323YesLysed whole bloodView Products
CD27M-T271YesLysed whole blood, lymphocytesView DataView Products
CD28CD28.2YesLysed whole bloodView Products
CD31WM59YesLysed whole bloodView Products
CD34581NoLysed whole bloodView Products
CD38HIT2YesLysed whole bloodView Products
CD405C3YesLysed whole bloodView Products
CD45HI30YesLysed whole bloodView Products
CD47CC2C6YesLysed whole blood, lymphocytesView DataView Products
CD50CBR-IC3/1YesLysed whole bloodView Products
CD56MEM-188YesLysed whole blood, lymphocytesView DataView Products
CD56HCD56YesLysed whole blood, lymphocytesView DataView Products
CD62LDREG-56YesLysed whole blood, lymphocytesView DataView Products
CD86IT2.2YesLysed whole blood, monocytesView DataView Products
CD88S5/1YesLysed whole blood, granulocytesView DataView Products
CD95DX2YesLysed whole blood, lymphocytesView DataView Products
CD99HCD99YesLysed whole blood, lymphocytesView DataView Products
CD127 (IL-7Rα)A019D5No Lysed whole blood, lymphocytesView DataView Products
CD134Ber-ACT35 (ACT35)NoLysed whole bloodView Products
CD172a/bSE5A5YesLysed whole bloodView Products
CD172gLSB2.20YesLysed whole bloodView Products
CD181 (CXCR1)8F1/CXCR1No Lysed whole blood, granulocytes. Lymphocytes and monocytes display high non-specific binding.View DataView Products
CD182 (CXCR2)5E8/CXCR2NoLysed whole blood, granulocytesView DataView Products
CD183 (CXCR3)G025H7NoLysed whole blood, lymphocytesView DataView Products
CD184 (CXCR4)12G5YesLysed whole blood, lymphocytesView DataView Products
CD193 (CCR3)5E8NoLysed whole blood, granulocytesView DataView Products
CD194 (CCR4)L291H4NoLysed whole blood, lymphocytesView DataView Products
CD195 (CCR5)T21/8NoLysed whole blood, lymphocytesView DataView Products
CD195 (CCR5)HEK/1/85aNoLysed whole blood, lymphocytesView DataView Products
CD195 (CCR5)J418F1NoLysed whole blood, lymphocytesView DataView Products
CD196 (CCR6)G034E3NoLysed whole blood, lymphocytesView DataView Products
CD197 (CCR7)G043H7NoLysed whole blood, lymphocytesView DataView Products
CD199 (CCR9)L053E8NoLysed whole blood, lymphocytesView DataView Products
CD244 (2B4)C1.7YesLysed whole bloodView Products
CD268 (BAFF-R)11C1YesLysed whole bloodView Products
CD279EH12.2H7NoLysed whole blood, lymphocytesView DataView Products
CD282 (TLR2)TL2.1YesLysed whole bloodView Products
CX3CR12A9-1NoLysed whole blood, lymphocytesView DataView Products
CX3CR1K0124E1NoLysed whole blood, lymphocytesView DataView Products
HLA-DRL243YesLysed whole bloodView Products
IgMMHM-88NoLysed whole bloodView Products

Chemokine receptors often display high false positives across lymphocytes, monocytes, and granulocytes when staining post-fixation.

Lymphocyte, monocyte, and granulocyte gates are based on FSC and SSC profiles.

FL-1 indicates the FITC Channel

Get your fixin’s, and learn more about our solutions for fixation:

True-Phos™ Perm Buffer is specially designed for the flow cytometric staining of phospho-specific targets like pSTAT3.

Fixation Buffer – for use with Permeabilization Wash Buffer (10X) to detect intracellular targets such as cytokines, chemokines, or other cytoplasmic antigens.

FluoroFix™ Buffer – for fixation of immunofluorescence stained cells, optimized to stabilize tandem dyes.

True-Nuclear™ Transcription Factor Buffer Set - for optimal staining of intranuclear and cytoplasmic antigens. Cytokine staining may not be optimal with this buffer as loss from cells will be evident with this method of fixation and permeabilization.  Refer to our Fixation and Permeabilization buffers above for cytokine staining.

RBC Lysis/Fixation Solution (10X) - optimized for one-step lysing of red blood cells (RBCs) and fixation of the remaining blood cells following immunofluorescent staining with fluorochrome-conjugated antibodies, suitable for either lyse/wash or lyse/no wash staining procedures.

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