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Flow cytometry analysis of V5 tag was done on HEK-293 cells transiently overexpressing V5-His-LacZ. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were labeled with V5 tag-FITC Mouse Monoclonal Antibody (R96325, red histogram) or with mouse isotype control (pink histogram) at a dilution of 1:250 in 2.5% BSA. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.
|Tested species reactivity||Tag|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||V5 synthetic peptide: Gly-Lys-Pro-Ile-Pro-Asn-Pro-Leu-Leu-Gly-Leu-Asp-Ser-Thr-.|
|Contains||0.01% sodium azide|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100-1:500|
|Western Blot (WB)||1:1000-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunocytochemistry (ICC)||See 3 publications below|
R963-25 recognizes amino acid sequence: -Gly-Lys-Pro-Ile-Pro-Asn-Pro-Leu-Leu-Gly-Leu-Asp-Ser-Thr-.
This antibody has been tested in IF experiments using cultured CHO (Chinese Hamster ovary) cells expressing recombinant epitope-tagged fusion proteins. For IF, dilute the FITC-conjugated antibody in PBS containing 10% fetal bovine serum (FBS). The amount of antibody provided is sufficient for 25 immunostaining reactions using a 1 mL working solution per reaction.
This antibody has also been tested against purified Positope™ control protein. The Positope™ control protein is a 53 kDa recombinant protein that contains seven epitope tags, including His(C-term), HisG, c-myc, and V5.
Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. The V5 epitope tag is derived from a small epitope (Pk) present on the P and V proteins of the paramyxovirus of simian virus (SV5). The V5 tag is usually used with all 14 amino acids (GKPIPNPLLGLDST), although it has also been used with a shorter 9 amino acid sequence (IPNPLLGLD).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Dendritic cells control fibroblastic reticular network tension and lymph node expansion.
R963-25 was used in immunocytochemistry to report that CLEC-2 modulation of podoplanin signaling permits fibroblastic reticular cell networks to stretch, which allows for rapid lymph node expansion
|Acton SE,Farrugia AJ,Astarita JL,Mourão-Sá D,Jenkins RP,Nye E,Hooper S,van Blijswijk J,Rogers NC,Snelgrove KJ,Rosewell I,Moita LF,Stamp G,Turley SJ,Sahai E,Reis e Sousa C||Nature (514:498)||2014|
|Not Applicable||Not Cited||
Evidence that bovine forebrain embryonic zinc finger-like gene influences immune response associated with mastitis resistance.
R963-25 was used in immunocytochemistry to study the role of FEZL-SEMA5A pathway in neuronal development and innate immunity in cows
|Sugimoto M,Fujikawa A,Womack JE,Sugimoto Y||Proceedings of the National Academy of Sciences of the United States of America (103:6454)||2006|
MLL 5 protein forms intranuclear foci, and overexpression inhibits cell cycle progression.
R963-25 was used in immunocytochemistry to elucidate the contribution of each domain of MLL5 to chromatin remodeling and cellular growth suppression
|Deng LW,Chiu I,Strominger JL||Proceedings of the National Academy of Sciences of the United States of America (101:757)||2004|
V5 Epitope Tag