Kinetics buffer

Name: Kinetics buffer
Brand: Sartorius
SKU: ESviCZIBPBHhf-iFVRba
Availability: InStock

Manufactured by Sartorius

34 citations

Sourced in Germany, United States

About the product

Kinetics buffer is a liquid buffer solution designed to maintain a stable pH environment for biochemical experiments and assays. It is formulated to provide consistent and reliable pH control during kinetic measurements and analyses.

Automatically generated - may contain errors

Get Technical Specifications Find protocols

Market Availability & Pricing

Is this product still available?

Get pricing insights and sourcing options

Lab products found in correlation

Check compatibility

34 protocols using «kinetics buffer»

Bifunctional Molecule Kinetics via Octet

2025

All Octet Assays were conducted using an Octet Red96e System (Sartorius). A volume of 200 μL of each solution was loaded onto a black flat-bottom 96-well plate (Grenier). Probes were always pre-wet for 10 minutes in 1X Kinetics Buffer (Sartorius) prior to the experiment. For ^uPARcGM kinetics, 300 nM of ^uPARcGM was incubated with 75 nM uPAR-Biotin from Acro Biosystems (Cat. No. UPR-H82E7) for various amounts of time. At the end of the incubation time a final concentration of 10 μM of AE105 uPAR competitor was used to quench the reaction. Streptavidin probes were placed in 1X Kinetics Buffer for 2 minutes to establish a baseline. This was followed by placing the probes in the timed incubation solution above, for 3 minutes. The probes were placed in 1X Kinetics Buffer for 3 minutes in order to re-establish a baseline. Next, they were added to a 250 nM Anti-DNP solution in 1X Kinetics Buffer for 5 minutes to measure the association. For ^AbcBM kinetics, 300 nM of ^AbcBM was incubated with 75 nM of Anti-DNP Ab for various amounts of time. 200 μM of DNP-Glycine competitor was used to quench the reaction. ProG probes (Sartorius) were placed in 1X Kinetics Buffer for 2 minutes to establish a baseline. This was followed by submerging the probes in the timed incubation solution above, for 3 minutes. The probes were then placed in 1X Kinetics Buffer for 3 minutes to re-establish a baseline. Next, they were added to the 500 nM uPAR solution in 1X Kinetics Buffer for 5 minutes to measure the association. For ^uPARcBM derivative kinetics, the above experiment was repeated, except the incubation time was 2 hours. For ^AbcBM terminal protein selectivity, 250 nM ^AbcBM was incubated with 100 nM uPAR-biotin overnight. Streptavidin probes were placed in 1X Kinetics Buffer for 2 minutes to establish a baseline. This was followed by placing the probes in the incubated solution above, for 3 minutes. The probes were placed in 1X Kinetics Buffer for 3 minutes in order to re-establish a baseline. Next, they were added to a 250 nM anti-DNP solution to measure association.
For ^AbcBM terminal protein selectivity, ^uPARcBM and ^uPARCBM-2 was incubated with 250 nM Ab overnight. The next day 200 μM DNP-glycine competitor was added to quench the reaction and disrupt any non-covalent complex. Streptavidin probes were placed in 200 μL of 1X Kinetics Buffer, to pre-wet for ten minutes. Next, the probes were placed in 1X Kinetics Buffer for 2 minutes to establish a baseline. This was followed by submerging the probes in a 100 nM solution of uPAR-Biotin from Acro Biosystems for 3 minutes. The probes were then placed in 1X Kinetics Buffer for 3 minutes in order to re-establish a baseline. Next, they were added to the 500 nM bifunctional molecule + 250 nM Anti-DNP + 200 μM DNP-Glycine solution in 1X Kinetics Buffer for 5 minutes to measure the association. For dual covalent reaction labeling, 100 nM bifunctional molecule was incubated with 100 nM uPAR biotin + 50 nM Anti-DNP overnight. The next day Streptavidin probes were placed in 1X Kinetics Buffer for 120 seconds to establish a baseline. This was followed by placing the probes in the incubated solutions above, for 3 minutes. The probes were then placed in 1X Kinetics Buffer with 10 μM AE105 competitor for 4 minutes. Next, they were added to a 50 μM DNP competitor solution for 5 minutes.

Kapcan E., Krygier K., da Luz M., Serniuck N.J., Zhang A., Bramson J, & Rullo A.F. (2025). Mimicry of molecular glues using dual covalent chimeras. Nature Communications, 16, 2855.

+ Open protocol

+ Expand

Affinity Analysis of Anti-CD28 Arm AI3

2025

Affinity of the S79 arm to PD-L1 was previously reported, whereas the affinity of the newly identified anti-CD28 arm AI3 was determined here by biolayer interferometry on an Octet RED96 instrument (Sartorius). All experiments were performed at 30°C. HIS1K biosensors (Sartorius) were loaded with CD28 (ACROBiosystems, Cat. #CD8-H52H3) at a concentration of 2 μg/mL for 300 seconds, and a loading signal of 0.5 nm was obtained for all experiments. A 2× serial dilution of NI-3201 was prepared in kinetics buffer (Sartorius), starting at 500 nmol/L, on seven different concentrations. The association and dissociation steps were monitored for 300 and 600 seconds, respectively. Data were processed and analyzed using Data Analysis Software (ForteBio). A double reference subtraction was applied, and both association and dissociation steps were fitted using a global 1.1 fitting model. For CD28 experiments, only the first 50 seconds of the dissociation phase were analyzed to improve fitting accuracy.

Majocchi S., Lloveras P., Nouveau L., Legrand M., Viandier A., Malinge P., Charreton M., Raymond C., Pace E.A., Millard B.L., Svensson L.A., Kelpšas V., Anceriz N., Salgado-Pires S., Daubeuf B., Magistrelli G., Gueneau F., Moine V., Masternak K., Shang L., Fischer N, & Ferlin W.G. (2025). NI-3201 Is a Bispecific Antibody Mediating PD-L1–Dependent CD28 Co-stimulation on T Cells for Enhanced Tumor Control. Cancer Immunology Research, 13(3), 365-383.

+ Open protocol

+ Expand

Aptamer Binding Kinetics on PIGF and VEGF

2024

The binding kinetics of the aptamers on PlGF and VEGF were evaluated using the Sartorius Octet^® R4 system manufactured by Sartorius, USA. Anti-penta HIS (HIS1K) biosensors purchased from Sartorius, USA, were used for measurements. Aptamer solutions were prepared using 1X Sartorius Kinetics Buffer purchased from Sartorius, USA. Prior to the experiment, the biosensors were hydrated in a pH 7.4 Phosphate buffer purchased from VWR, USA, for 10 min. An initial baseline step of 60 s was done in pH 7.4 phosphate buffer since the protein stock solutions were dissolved in that buffer, and the proteins were loaded on the biosensors by immersing the biosensors for 300 s in protein solutions. PlGF and VEGF loading concentrations were kept at 2.5 µg/mL and 10 µg/mL respectively, since a significant binding response could be observed at those concentrations. It was followed by a secondary baseline step of 60 s and association and dissociation steps for 300 s each. The association and dissociation profiles of the aptamers were determined at concentrations of 0, 2.5, 5, 10, and 20 µM. A set of non-loaded reference sensors was used to mitigate the impact of non-specific binding. Binding parameters were calculated by Octet ^® Analysis Studio software using a 1:1 global-fitting model. All experiments were carried out in triplicate.

Mallawarachchi S., Cebecioglu R.E., Althumayri M., Beker L., Fernando S, & Ceylan Koydemir H. (2024). Systematic design and evaluation of aptamers for VEGF and PlGF biomarkers of Preeclampsia. BMC Biotechnology, 24, 64.

+ Open protocol

+ Expand

Kinetic Analysis of Antibody-Antigen Interactions

2024

For affinity determination, anti-human IgG-Fc capture (AHC, Sartorius, Göttingen, Germany) biosensors were equilibrated in PBS for 10 min and loaded with 10 µg/mL antibody solution until a layer thickness of 1 nm was reached. After a 30 s quenching step in kinetics buffer (KB, Sartorius), association was measured for 300 s against varying antigen concentrations diluted in KB. All antigens were produced in-house (B7-H3 with C-terminal StrepII-Tag, TIGIT and CD16 with N-terminal His₆ and C-terminal TwinStrep-Tag), and analyzed concentrations ranged from 500 nM to 7.8 nM. Subsequent dissociation in KB was measured for 300 s. KB without antigen was used as a negative control. Binding kinetics were calculated based on Savitzky-Golay filtering and a 1:1 Langmuir binding model.
For the simultaneous binding assay, AHC biosensors were equilibrated in PBS for 10 min and loaded with 10 µg/mL antibody until a layer thickness of 1 nm was reached. After a 30 s quenching step in PBS, association was measured sequentially for a 250 nM solution of all antigens. As a control, the association of only one or two antigens with subsequent incubation in PBS was also measured. The antigens used were B7-H3-SII, TIGIT, and His-CD16-TwinStrep.
All measurements were performed using the Octet RED96 system (ForteBio, Fremont, CA, USA) at 30 °C and 1000 rpm.

Ulitzka M., Harwardt J., Lipinski B., Tran H., Hock B, & Kolmar H. (2024). Potent Apoptosis Induction by a Novel Trispecific B7-H3xCD16xTIGIT 2+1 Common Light Chain Natural Killer Cell Engager. Molecules, 29(5), 1140.

+ Open protocol

+ Expand

Corresponding organizations : Technical University of Darmstadt

Kinetic analysis of anti-TcPOP mAb binding

2024

Binding of anti-TcPOP mAbs to TcPOP were measured by kinetic experiments carried out on an Octet R4 (Sartorius). All samples were buffer exchanged into Sartorius Kinetics Buffer, according to the manufacturer's instructions. All measurements were performed at 200 μl/well in Sartorius kinetic buffer at 25°C in 96-well black plates (Greiner Bio-One, Cat# 655209). ProG (Cat. Nos. 18-5082, 18-5083, 18-5084) were used to immobilize anti-TcPOP mAbs for 1800s. Immunogens were four-fold serially diluted in kinetic buffer in the range of 64 nM to 4 nM. Assay was performed in three sequential steps with Octet BLI Discovery 12.2.2.20 software (Sartorius):
Step 1, biosensor hydration and equilibration (300s); Step 2, immobilization of anti-TcPOP IgG1 mAbs on a ProG biosensor (600s);
Step 3, wash and establish baseline (60s); Step 4, measure TcPOP association kinetics (1800s); and
Step 5, measure TcPOP dissociation kinetics (600s). The acquired raw data for the binding of anti-TcPOP mAbs with TcPOP were processed and globally fit to a 1:1 binding model. Binding kinetics measurements were conducted in triplicate, and reported values represent the average. Data were analysed using Octet Analysis Studio 12.2.2.26 Software (Sartorius) and graphs produced using GraphPad (Prism).

Batra S., Ragan T.J., Frank A.M., Kaplan M., Lancey C., Assadipapari M., Ying C., Struwe W.B., Hesketh E.L., Barfod L., & Campeotto I. (2024). Open and closed conformations of a sub-80 kDa Chagas vaccine candidate defined by a cryo-EM led integrative approach.

+ Open protocol

+ Expand

Corresponding organizations : University of Nottingham, University of Leicester, University of Copenhagen, University of Oxford, Nottingham Trent University

Top 5 most cited protocols using «kinetics buffer»

Monovalent and Avidity Fab Binding Assays

Cited 1 time

Bio-layer interferometry measurements were performed using a ForteBio Octet Red96 (Sartorius). Monovalent binding assays were performed using High precision streptavidin biosensors (Sartorius), loaded with ¹⁶NIP-BSA-biotin (Biosearch Technologies) [5.86nM]. Fabs were diluted in 1x Kinetics Buffer (KB) (Sartorius) and assayed at 100, 50, and 25 nM. Ligand-coated biosensors were regenerated by short incubation in HCl (Sigma-Aldrich) buffer followed by neutralization in 1x KB. For avidity measurements, Anti-human Fab-CH1 biosensors (Sartorius) were loaded with Fabs diluted in 1x KB [100 nM] and assayed with either ²NP-BSA or ⁹NP-BSA (Biosearch Technologies) at 0.33 and 0.11 μM.

Otto-Bruc A.E., Fariss R.N., Van Hooser J.P, & Palczewski K. (1998). Phosphorylation of photolyzed rhodopsin is calcium-insensitive in retina permeabilized by α-toxin. Proceedings of the National Academy of Sciences of the United States of America, 95(25), 15014-15019.

+ Open protocol

+ Expand

Corresponding organizations : Rockefeller University, Howard Hughes Medical Institute

Characterizing Chimeric Antibody Interactions

Cited 1 time

For affinity determination of chimeric antibodies, anti-human IgG-Fc capture (AHC) biosensors were equilibrated in PBS pH 7.4 for 10 min and subsequently loaded with 10 µg/ml of the antibody of interest until a layer thickness of 1 nm was reached. All following steps were performed using kinetics buffer (KB, Sartorius). Association was measured for 600 s using varying concentrations of EGFR-ECD or PD-L1-ECD (produced in-house) ranging from 7.8 nM to 500 nM followed by dissociation for 600 s. KB was used as a negative control. Binding kinetics were determined based on Savitzky-Golay filtering and a 1:1 Langmuir binding model.
For the EGF competition assay, AHC biosensors were loaded with 10 µg/ml of HCP-LCE until a layer thickness of 1 nm was reached. Subsequently, 250 nM EGFR-ECD pre-incubated with either 0 nM, 250 nM or 1000 nM EGF was applied for 600 s.
For the PD-1 competition assay, anti-human Fab-CH₁ 2nd Generation (FAB2G) biosensors were loaded with 10 µg/ml of HCP-LCE until a layer thickness of 1 nm was reached. Subsequently, 250 nM PD-L1-ECD pre-incubated with either 0 nM, 250 nM or 1000 nM PD-1 was applied for 600 s.
For the simultaneous binding assay, AHC biosensors were loaded with 10 µg/ml of oaHCP-LCE until a layer thickness of 1 nm was reached. After measurement of the association to 250 nM antigen 1 for 300 s, association to 250 nM antigen 2 was determined for 300 s. As controls, oaHCP-LCE was incubated with antigen 1 or PBS only. EGFR-ECD and PD-L1-ECD were used as antigens and the order of association was analyzed in both settings.
All measurements were performed using the Octet RED96 system (FortéBio, Molecular Devices) at 30°C and 1000 rpm.

Harwardt J., Bogen J.P., Carrara S.C., Ulitzka M., Grzeschik J., Hock B, & Kolmar H. (2022). A Generic Strategy to Generate Bifunctional Two-in-One Antibodies by Chicken Immunization. Frontiers in Immunology, 13, 888838.

+ Open protocol

+ Expand

Corresponding organizations : Technical University of Darmstadt, Ferring Pharmaceuticals (Germany), Ferring Pharmaceuticals (Switzerland)

Biosensor-Based Kinetic Analysis of Omicron RBD Binding

BLI experiments were performed using an Octet R8 system (Sartorius). Biotinylated SARS-CoV-2 Omicron RBDs (ACRObiosystems) or 6x(His)-tagged SARS-CoV-1 RBD (R&D Systems) at 5 nM was loaded onto streptavidin or Ni-NTA biosensor tips (Sartorius), respectively, immersed in Kinetics Buffer (Sartorius) for 120 s. For measuring K_D, the antibodies at five different concentrations were subjected to BLI runs with an association step (for 240 or 360 s) and a dissociation step (for 720 s). The binding kinetics were analyzed with the Octet BLI Analysis 12.2.2.4 software package (Sartorius).

Jeong B.S., Jeon J.Y., Lai C.J., Yun H.Y., Jung J.U, & Oh B.H. (2022). Structural basis for the broad and potent cross-reactivity of an N501Y-centric antibody against sarbecoviruses. Frontiers in Immunology, 13, 1049867.

+ Open protocol

+ Expand

Corresponding organizations : Korea Advanced Institute of Science and Technology, Cleveland Clinic Lerner College of Medicine

Biolayer Interferometry of GPIbα-VWF Binding

BLI experiments were performed using an Octet QK^e instrument with data acquired at 0.3 Hz with 1000 rpm shaking during all steps. Samples were diluted in kinetics buffer (ForteBio/Sartorius), comprised of PBS, 0.1% BSA, and 0.02% Tween-20. Equilibrated streptavidin (SA) sensors were loaded with ~10 μg/mL GPIbα-LBD from either species until hitting a set threshold for 2 nm of binding. Sensors were then washed briefly and set to association in various VWF fragments for 100 seconds and dissociate for 500 seconds. Loaded sensors were washed with 1 M NaCl to remove any remaining bound VWF and equilibrated again for the next samples. BLI experiments were repeated at least twice for each binding interaction. Sensorgrams presented are representative samples. Data was fit to a heterogenous ligand binding model using two rate equations. Kinetic parameters can be found in Supplemental Table 3. To obtain equilibrium response (ΣR_eq K_D) values for each binding reaction, Req1 and Req2 were summed for each concentration and fitted to a hyperbola assuming a 1:1 stoichiometric binding ratio.

Arce N.A., Liu Y., Chen W., Zhang X.F, & Li R. (2022). Autoinhibitory module underlies species difference in shear activation of von Willebrand factor. Journal of thrombosis and haemostasis : JTH, 20(11), 2686-2696.

+ Open protocol

+ Expand

Corresponding organizations : Children's Healthcare of Atlanta, Emory University, Aflac (United States), Lehigh University

TfR Binding Assay by BLI

Competition of ETV:IDS and IgG:IDS for hTfR binding was assessed using biolayer interferometry. Biolayer interferometry experiments were performed on an Octet RED384 (Sartorius) instrument at 25°C. All the samples were diluted with Kinetics buffer (18-1092; Sartorius) and transferred to solid-black 384-well plates (781900; Greiner Bio-One). Streptavidin-coated biosensors (18-5021; Sartorius) were loaded with biotinylated recombinant human TfR ECD for 400 s. TfR binding antibody, IgG:IDS, and ETV:IDS were subsequently loaded for 1,100 s on surface-bound TfR followed by ETV:IDS loading for hTfR-binding competition.

Arguello A., Mahon C.S., Calvert M.E., Chan D., Dugas J.C., Pizzo M.E., Thomsen E.R., Chau R., Damo L.A., Duque J., Fang M., Giese T., Kim D.J., Liang N., Nguyen H.N., Solanoy H., Tsogtbaatar B., Ullman J.C., Wang J., Dennis M.S., Diaz D., Gunasekaran K., Henne K.R., Lewcock J.W., Sanchez P.E., Troyer M.D., Harris J.M., Scearce-Levie K., Shan L., Watts R.J., Thorne R.G., Henry A.G, & Kariolis M.S. (2022). Molecular architecture determines brain delivery of a transferrin receptor–targeted lysosomal enzyme. The Journal of Experimental Medicine, 219(3), e20211057.

+ Open protocol

+ Expand

Corresponding organizations : Denali Therapeutics (United States), University of Minnesota

Spelling variants (same manufacturer)

Octet Kinetics Buffer - 22 citations 1X kinetic buffer - 8 citations

Similar products (other manufacturers)

Kinetics buffer (by Molecular Devices) - 112 citations Kinetic buffer (by Molecular Devices) - 41 citations Octet Kinetics buffer (by Molecular Devices) - 7 citations Kinetics (by MathWorks) - 6 citations Kinetics (by FTS Systems) - 4 citations PBS Kinetics Buffer (by Molecular Devices) - 3 citations Streptavidin biosensors and Kinetics Buffer (by Molecular Devices) - 3 citations Kinetic (by Molecular Devices) - 2 citations Kinetic (by Helena Chemical) - 2 citations BLI Kinetic buffer (by Molecular Devices) - 2 citations

The spelling variants listed above correspond to different ways the product may be referred to in scientific literature.
These variants have been automatically detected by our extraction engine, which groups similar formulations based on semantic similarity.

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?

Revolutionizing how scientists
search and build protocols!