Log In Sign up
The largest database of trusted experimental protocols

Sf buffer

Manufactured by Lonza
Visit Supplier

SF buffer is a laboratory reagent used to maintain a specific pH during various biological and biochemical experiments. It is a solution that provides a stable buffering environment for such activities.

Automatically generated - may contain errors

Lab products found in correlation

100 μl cuvette, by Lonza (4 mentions) Amaxa 4d nucleofector system, by Lonza (4 mentions) Nucleofector 96 well shuttle system, by Lonza (2 mentions) Paromomycin, by Biosynth (2 mentions) Papain, by Merck Group (2 mentions) Synthetic sgrna, by Synthego (2 mentions) Tryple express, by Thermo Fisher Scientific (2 mentions) 4d nucleofector, by Lonza (2 mentions) Ze5 cell analyzer, by Bio-Rad (1 mentions) Lightning link r pe conjugation kit, by Abcam (1 mentions) Mem non essential amino acid, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin glutamine (psg), by Thermo Fisher Scientific (1 mentions) Bhk 21 cells, by American Type Culture Collection (1 mentions) Dual luciferase assay, by Promega (1 mentions) Se buffer, by Lonza (1 mentions) 4d nucleofector system, by Lonza (1 mentions) Paromomycin, by Merck Group (1 mentions) Amaxa 4d nucleofector, by Lonza (1 mentions) Electroporator, by Lonza (1 mentions) Bovine serum albumin fraction 5, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

SF nucleofection buffer (3 citations) SF (K562) or SG (Raji and Ramos) buffer (2 citations) SE/SF/SG buffer (2 citations) SF electroporation buffer (2 citations) Cell line buffer SF (2 citations) 20 µl SF buffer (2 citations)

19 protocols using sf buffer

1

Quantification of dsRNA in Electroporated BHK-21 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
BHK-21 cells [American Type Culture Collection (ATCC), Manassas, VA, CCL10] were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS; Gemini, #100-106), MEM nonessential amino acids (Gibco, #11140050), and penicillin/streptomycin/glutamine (Gibco, #10378016). BHK-21 cells were electroporated in strip cuvettes with 3 μg of RNA per 106 cells using SF buffer (Lonza) and 4D-Nucleofector. At 20 hours after electroporation, cells were removed from wells by incubation with trypsin-EDTA solution and washed in PBS containing 5% bovine serum albumin (BSA). The cells were stained for dsRNA using the J2 anti-dsRNA Ab (Scicons, #10010500) conjugated to R-PE using a Lightning-Link R-PE conjugation kit (Innova Biosciences). After staining, cells were evaluated on a ZE5 Cell analyzer (Bio-Rad) and the data were analyzed using FlowJo 10 (Tree Star, Ashland, OR).
Elong Ngono A., Syed T., Nguyen A.V., Regla-Nava J.A., Susantono M., Spasova D., Aguilar A., West M., Sparks J., Gonzalez A., Branche E., DeHart J.L., Vega J.B., Karmali P.P., Chivukula P., Kamrud K., Aliahmad P., Wang N, & Shresta S. (2020). CD8+ T cells mediate protection against Zika virus induced by an NS3-based vaccine. Science Advances, 6(45), eabb2154.
+ Open protocol
+ Expand
2

BCL6 Silencing in DLBCL Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
DLBCL cells (3 × 106 to 5 × 106) were transfected with 1 micromolar siRNA for BCL6 (siBCL6-1 HSS100968 Life Technologies, siBCL6-2 SI00311129 Qiagen, siBCL6-3 SI0031143 Qiagen and siBCL6-4 SI0031150 Qiagen) or control (GFP targeting siRNA) using 96-well electroporation (SF buffer, Lonza).
Dupont T., Yang S., Patel J., Hatzi K., Malik A., Tam W., Martin P., Leonard J., Melnick A, & Cerchietti L. (2015). Selective targeting of BCL6 induces oncogene addiction switching to BCL2 in B-cell lymphoma. Oncotarget, 7(3), 3520-3532.
+ Open protocol
+ Expand
3

Genetic Modification of 293T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
293T cells were transduced at a low MOI (<0.3) with a lentivirus expressing BFP under the control of an EF1α promoter (Addgene #71825) and sorted by flow cytometry to produce a pure population of BFP-expressing cells. One day post-AAV transduction, 2 × 105 cells were resuspended in 20 µl SF buffer (Lonza) and nucleofected with 10 µl sgBFP RNP and 100 pmol of ssODN donor, using program DS-150 on a Nucleofector 96-well Shuttle system (Lonza). Cells were then transferred to a 12-well plate containing pre-warmed medium, and grown for 4 days. BFP and GFP fluorescence were measured by flow cytometry on a BD Fortessa, and the results were analyzed using FlowJo v10 software.
Canny M.D., Moatti N., Wan L.C., Fradet-Turcotte A., Krasner D., Mateos-Gomez P.A., Zimmermann M., Orthwein A., Juang Y.C., Zhang W., Noordermeer S.M., Seclen E., Wilson M.D., Vorobyov A., Munro M., Ernst A., Ng T.F., Cho T., Cannon P.M., Sidhu S.S., Sicheri F, & Durocher D. (2017). Inhibition of 53BP1 favors homology-dependent DNA repair and increases CRISPR-Cas9 genome-editing efficiency. Nature biotechnology, 36(1), 95-102.
+ Open protocol
+ Expand
4

Genetic Modification of Plasmodium Sporozoites

Check if the same lab product or an alternative is used in the 5 most similar protocols
Oocysts (1.25 × 107 per transfection) were excysted as described above, and sporozoites were pelleted by centrifugation and resuspended in SF buffer (Lonza) containing 50 μg of tagging or targeting plasmids and 30 μg CRISPR/Cas9 plasmid in a total volume of 100 μl. The mixtures were then transferred to a 100-μl cuvette (Lonza) and electroporated on an AMAXA 4D-Nucleofector system (Lonza) using program EH100. Electroporated sporozoites were transferred to cold DPBS and kept on ice before infecting mice.
Xu R., Feng Y., Xiao L, & Sibley L.D. (2021). Insulinase-like Protease 1 Contributes to Macrogamont Formation in Cryptosporidium parvum. mBio, 12(2), e03405-20.
+ Open protocol
+ Expand
5

Targeted Genomic Editing by sRGN3.1 and T428

Check if the same lab product or an alternative is used in the 5 most similar protocols
Forty-nine sites containing up to five nucleotide substitutions from the T428 target sites were selected for analysis. The homozygous IVS40 293FT cell line (2 × 105 cells) was transfected with 2 µg of a plasmid carrying sRGN3.1 and T428 sgRNA in Lonza SF buffer by nucleofection under Program CM-130, and genomic DNA was extracted 7 days after nucleofection. All the selected sites except one on-target site were successfully PCR-amplified, and indels were analyzed by NGS as described in the section Amplicon sequencing (AmpSeq) and NGS analysis.
Schmidt M.J., Gupta A., Bednarski C., Gehrig-Giannini S., Richter F., Pitzler C., Gamalinda M., Galonska C., Takeuchi R., Wang K., Reiss C., Dehne K., Lukason M.J., Noma A., Park-Windhol C., Allocca M., Kantardzhieva A., Sane S., Kosakowska K., Cafferty B., Tebbe J., Spencer S.J., Munzer S., Cheng C.J., Scaria A., Scharenberg A.M., Cohnen A, & Coco W.M. (2021). Improved CRISPR genome editing using small highly active and specific engineered RNA-guided nucleases. Nature Communications, 12, 4219.
+ Open protocol
+ Expand
6

Genetic Modification of 293T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
293T cells were transduced at a low MOI (<0.3) with a lentivirus expressing BFP under the control of an EF1α promoter (Addgene #71825) and sorted by flow cytometry to produce a pure population of BFP-expressing cells. One day post-AAV transduction, 2 × 105 cells were resuspended in 20 µl SF buffer (Lonza) and nucleofected with 10 µl sgBFP RNP and 100 pmol of ssODN donor, using program DS-150 on a Nucleofector 96-well Shuttle system (Lonza). Cells were then transferred to a 12-well plate containing pre-warmed medium, and grown for 4 days. BFP and GFP fluorescence were measured by flow cytometry on a BD Fortessa, and the results were analyzed using FlowJo v10 software.
Canny M.D., Moatti N., Wan L.C., Fradet-Turcotte A., Krasner D., Mateos-Gomez P.A., Zimmermann M., Orthwein A., Juang Y.C., Zhang W., Noordermeer S.M., Seclen E., Wilson M.D., Vorobyov A., Munro M., Ernst A., Ng T.F., Cho T., Cannon P.M., Sidhu S.S., Sicheri F, & Durocher D. (2017). Inhibition of 53BP1 favors homology-dependent DNA repair and increases CRISPR-Cas9 genome-editing efficiency. Nature biotechnology, 36(1), 95-102.
+ Open protocol
+ Expand
7

Genetic Modification of Malaria Parasites

Check if the same lab product or an alternative is used in the 5 most similar protocols
The 1 × 108 sporozoites were resuspended in SF buffer (Lonza) containing 50 μg of tagging or knockout plasmids, or 30 μg amplicon DNA, together with 30 μg CRISPR/Cas9 plasmid in a total 200 μl volume. The mixtures were then transferred to a 100-μl cuvette (Lonza) and electroporated on an AMAXA 4D-Nucleofector system (Lonza) using program EH100. Electroporated sporozoites were transferred to cold DPBS and kept on ice before infection. To select transgenic parasites, GKO mice were orally given 8% (wt/vol) sodium bicarbonate 5 min prior infection and then orally gavaged with electroporated sporozoites. Transgenic parasites were selected with 16 g/l paromomycin (Biosynth International, Inc) provided in the drinking water starting at 24 h and continuing ad lib. To amplify transgenic parasites or monitor oocysts burden, NSG mice were administered the fecal slurry containing 2 × 104 oocysts from an infected GKO mouse, and paromomycin drinking water was used for selection immediately after infection. Fecal pellets were collected after 3 dpi and stored at −80 °C for DNA extraction or at 4 °C for luciferase assay or for purification.
Xu R., Beatty W.L., Greigert V., Witola W.H, & Sibley L.D. (2023). Multiple pathways for glucose phosphate transport and utilization support growth of Cryptosporidium parvum. bioRxiv.
+ Open protocol
+ Expand
8

Electroporation of ZFL Cells for IFNϕ1 Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
ZFL cells were grown to 80% confluency prior to transfection. Cells were washed, treated with trypsin, centrifuged at 90×g for 10 min and resuspended to a final concentration of 2×107 cells/mL in SF buffer (Lonza). Using the 96-well transfection module (Lonza), a total of 500ng DNA was added to 4×105 cells in 20µL and the cells were electroporated using the program EW-158. Cells were then resuspended in 180µL of LDF media in quadruplicate and allowed to recover for 24 h. For reporter assays ZFL cells we re transiently transfected as described above with frm expression vector containing mda5, ΔCARDmda5, or empty vector (250ng) along with 250ng of an IFNϕ1 promoter-driven luciferase reporter (pGL3-IFN) (Sullivan et al., 2007 ) and the Renilla-expressing internal control pRL-CMV (6.25ng) (Promega). After cells were allowed to recover for 24 h , the medium was removed and cells were exposed to SHRV at MOI = 0.1 in MEM for 1 h followed by supplementation with LDF. Dual-luciferase assays (Promega) were performed in triplicate 24 h post infection.
Gabor K., Charette J., Pietraszewski M., Wingfield D., Shim J., Millard P, & Kim C. (2015). A DN-mda5 Transgenic Zebrafish 1 Model Demonstrates that Mda5 Plays an Important Role in Snakehead Rhabdovirus Resistance. Developmental and comparative immunology, 51(2), 298-304.
+ Open protocol
+ Expand
9

Priming S. rosetta Cells for Nucleofection

Check if the same lab product or an alternative is used in the 5 most similar protocols
To prime S. rosetta cells for nucleofection, we treated them with a cocktail that removes the extracellular matrix as follows. Aliquots of washed cells were pelleted at 800 g and 22°C for 5 min. The supernatant was gently removed with gel-loading tips and each pellet was resuspended in 100 µl of priming buffer (40 mM HEPES-KOH, pH 7.5; 34 mM lithium citrate; 50 mM l-cysteine; 15% [wt/vol] PEG 8000; and 1 μM papain [Millipore Sigma, St. Louis, MO; Cat. No. P3125-100MG]). After incubating cells for 30–40 min, 10 µl of 50 mg/ml bovine serum albumin was added to each aliquot of primed cells to quench proteolysis from the priming buffer. Finally, the cells were centrifuged at 1250 g and 22°C for 5 min, the supernatant was removed, and the pellet was resuspended in 25 µl of SF Buffer (Lonza, Basel, Switzerland; Cat. No. V4SC-2960). The resuspended cells were stored on ice while preparing nucleofection reagents.
Booth D.S, & King N. (2020). Genome editing enables reverse genetics of multicellular development in the choanoflagellate Salpingoeca rosetta. eLife, 9, e56193.
+ Open protocol
+ Expand
10

Caco-2 Cell Transfection with SpCas9 RNP

Check if the same lab product or an alternative is used in the 5 most similar protocols
For Caco-2 transfection, 10 pmol Streptococcus Pyogenes NLS-Sp.Cas9-NLS (SpCas9) nuclease (Aldevron; 9212) was combined with 30 pmol total synthetic sgRNA (10 pmol each sgRNA, Synthego) to form ribonucleoproteins (RNPs) in 20 μl total volume with SF Buffer (Lonza V5SC-2002) and allowed to complex at room temperature for 10 min.
All cells were dissociated into single cells using TrypLE Express (Gibco), resuspended in culture media and counted. 100,000 cells per nucleofection reaction were pelleted by centrifugation at 200 × g for 5 min. After centrifugation, cells were resuspended in transfection buffer according to cell type and diluted to 2 × 104 cells/μl. Five μl of cell solution was added to preformed RNP solution and gently mixed. Nucleofections were performed on a Lonza HT 384-well nucleofector system (Lonza, #AAU-1001) using program CM-150 for Caco-2. Immediately after nucleofection, each reaction was transferred to a tissue-culture treated 96-well plate containing 100 μl of normal culture media and seeded at a density of 50,000 cells per well. Transfected cells were incubated following standard protocols.
Gordon D.E., Hiatt J., Bouhaddou M., Rezelj V.V., Ulferts S., Braberg H., Jureka A.S., Obernier K., Guo J.Z., Batra J., Kaake R.M., Weckstein A.R., Owens T.W., Gupta M., Pourmal S., Titus E.W., Cakir M., Soucheray M., McGregor M., Cakir Z., Jang G., O’Meara M.J., Tummino T.A., Zhang Z., Foussard H., Rojc A., Zhou Y., Kuchenov D., Hüttenhain R., Xu J., Eckhardt M., Swaney D.L., Fabius J.M., Ummadi M., Tutuncuoglu B., Rathore U., Modak M., Haas P., Haas K.M., Naing Z.Z., Pulido E.H., Shi Y., Barrio-Hernandez I., Memon D., Petsalaki E., Dunham A., Marrero M.C., Burke D., Koh C., Vallet T., Silvas J.A., Azumaya C.M., Billesbølle C., Brilot A.F., Campbell M.G., Diallo A., Dickinson M.S., Diwanji D., Herrera N., Hoppe N., Kratochvil H.T., Liu Y., Merz G.E., Moritz M., Nguyen H.C., Nowotny C., Puchades C., Rizo A.N., Schulze-Gahmen U., Smith A.M., Sun M., Young I.D., Zhao J., Asarnow D., Biel J., Bowen A., Braxton J.R., Chen J., Chio C.M., Chio U.S., Deshpande I., Doan L., Faust B., Flores S., Jin M., Kim K., Lam V.L., Li F., Li J., Li Y.L., Li Y., Liu X., Lo M., Lopez K.E., Melo A.A., Moss FR I.I.I., Nguyen P., Paulino J., Pawar K.I., Peters J.K., Pospiech TH J.r., Safari M., Sangwan S., Schaefer K., Thomas P.V., Thwin A.C., Trenker R., Tse E., Tsui T.K., Wang F., Whitis N., Yu Z., Zhang K., Zhang Y., Zhou F., Saltzberg D., Hodder A.J., Shun-Shion A.S., Williams D.M., White K.M., Rosales R., Kehrer T., Miorin L., Moreno E., Patel A.H., Rihn S., Khalid M.M., Vallejo-Gracia A., Fozouni P., Simoneau C.R., Roth T.L., Wu D., Karim M.A., Ghoussaini M., Dunham I., Berardi F., Weigang S., Chazal M., Park J., Logue J., McGrath M., Weston S., Haupt R., Hastie C.J., Elliott M., Brown F., Burness K.A., Reid E., Dorward M., Johnson C., Wilkinson S.G., Geyer A., Giesel D.M., Baillie C., Raggett S., Leech H., Toth R., Goodman N., Keough K.C., Lind A.L., Klesh R.J., Hemphill K.R., Carlson-Stevermer J., Oki J., Holden K., Maures T., Pollard K.S., Sali A., Agard D.A., Cheng Y., Fraser J.S., Frost A., Jura N., Kortemme T., Manglik A., Southworth D.R., Stroud R.M., Alessi D.R., Davies P., Frieman M.B., Ideker T., Abate C., Jouvenet N., Kochs G., Shoichet B., Ott M., Palmarini M., Shokat K.M., García-Sastre A., Rassen J.A., Grosse R., Rosenberg O.S., Verba K.A., Basler C.F., Vignuzzi M., Peden A.A., Beltrao P, & Krogan N.J. (2020). Comparative host-coronavirus protein interaction networks reveal pan-viral disease mechanisms. Science (New York, N.y.), 370(6521), eabe9403.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!