Highly sensitive detection of antibody nonspecific interactions using flow cytometry

The quickly evolving nature of antibody drug improvement has resulted in applied sciences that generate huge numbers (a whole bunch to hundreds) of lead antibody candidates throughout early discovery. These candidates have to be quickly pared all the way down to establish probably the most drug-like candidates for in-depth evaluation of their security and efficacy, which may solely be carried out on a restricted variety of antibodies attributable to time and useful resource necessities. One key biophysical property of profitable antibody therapeutics is excessive specificity, outlined as low ranges of nonspecific binding or polyspecificity.

Though there was some progress in creating assays for detecting antibody polyspecificity, most of those assays are restricted by poor sensitivity or assay codecs that require proprietary antibody floor show strategies, and a few of these assays use advanced and poorly outlined polyspecificity reagents. Right here we report the PolySpecificity Particle (PSP) assay, a delicate circulation cytometry assay for evaluating antibody nonspecific interactions that overcomes earlier limitations and can be utilized for evaluating various sorts of IgGs, multispecific antibodies and Fc-fusion proteins. Our strategy makes use of micron-sized magnetic beads coated with Protein A to seize antibodies at extraordinarily dilute concentrations (<0.02 mg/mL).

Move cytometry evaluation of polyspecificity reagent binding to those conjugates ends in delicate detection of variations in nonspecific interactions for clinical-stage antibodies. Our PSP assay strongly discriminates between antibodies with totally different ranges of polyspecificity utilizing beforehand reported polyspecificity reagents which are both well-defined proteins or extremely advanced protein mixtures. Furthermore, we additionally discover {that a} distinctive reagent, particularly ovalbumin, ends in the very best assay sensitivity and specificity. Importantly, our assay is far more delicate than customary assays equivalent to ELISAs. We anticipate that our easy, delicate, and high-throughput PSP assay will speed up the event of protected and efficient antibody therapeutics.

Toxicity of Carbon Nanomaterials-In the direction of Dependable Viability Evaluation by way of New Method in Move Cytometry

The scope of software of carbon nanomaterials in biomedical, environmental and industrial fields is lately considerably growing. Since in vitro toxicity testing is the primary important step for any business utilization, it’s essential to have a dependable methodology to investigate the possibly dangerous results of carbon nanomaterials. Regardless that researchers already reported the interference of carbon nanomaterials with widespread toxicity assays, there’s nonetheless, sadly, a lot of research that neglect this reality. On this research, we investigated interference of 4 bio-promising carbon nanomaterials (graphene acid (GA), cyanographene (GCN), graphitic carbon nitride (g-C3N4) and carbon dots (QCDs)) in generally used LIVE/DEAD assay.

When a regular process was utilized, supplies prompted varied sorts of interference. Whereas positively charged g-C3N4 and QCDs induced false outcomes by means of the creation of free agglomerates and intrinsic fluorescence properties, negatively charged GA and GCN led to false indicators as a result of advanced quenching impact of the fluorescent dye of a LIVE/DEAD package. Thus, we developed a brand new strategy utilizing a particular gating technique primarily based on extra controls that efficiently overcame all sorts of interference and result in dependable ends in LIVE/DEAD assay.

We advise that the newly developed process ought to be a compulsory device for all in vitro circulation cytometry assays of any class of carbon nanomaterials. Imaging circulation cytometry permits for the quantitative evaluation of fluorescent indicators on the subcellular degree. Right here, we describe using a biosensor cell line, particularly, U2OS osteosarcoma cells outfitted with a fusion protein comprising monomeric pink fluorescent protein (mRFP), inexperienced fluorescent protein (GFP) and microtubule-associated proteins 1A/1B gentle chain 3B (greatest often called LC3), for the evaluation of autophagic flux by imaging circulation cytometry.

Highly sensitive detection of antibody nonspecific interactions using flow cytometry

We element all evaluation instruments required to tell apart autophagosomes (that emit each a pink and a inexperienced fluorescence) and autolysosomes (that emit a pink fluorescence, but lose the inexperienced fluorescent sign) and to quantitate autophagic flux in a handy style. Our information, nevertheless, present that information of the kinetics of the interplay permits to reconcile the info obtained by circulation cytometry and LigandTracer and show the complementarity of those two strategies.

Okay D willpower from time-resolved experiments on stay cells with LigandTracer and reconciliation with end-point circulation cytometry measurements

Design of next-generation therapeutics comes with new challenges and emulates expertise and strategies to satisfy them. Characterizing the binding of both pure ligands or therapeutic proteins to cell-surface receptors, for which related recombinant variations could not exist, represents certainly one of these challenges. Right here we report the characterization of the interplay of 5 totally different antibody therapeutics (Trastuzumab, Rituximab, Panitumumab, Pertuzumab, and Cetuximab) with their cognate goal receptors utilizing LigandTracer.

Rabbit Polyclonal antibody Anti-CRBN

Anti-CRBN 50 µg
EUR 349

Anti-MCCC2 (2B3)

YF-MA19192 100 ug
EUR 363
Description: Mouse monoclonal to MCCC2

MCCC2 Antibody

1-CSB-PA013573GA01HU
  • EUR 597.00
  • EUR 333.00
  • 150ul
  • 50ul
Description: A polyclonal antibody against MCCC2. Recognizes MCCC2 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC

MCCC2 Antibody

1-CSB-PA867199ESR1HU
  • EUR 222.00
  • EUR 335.00
  • 100ul
  • 50ul
Description: A polyclonal antibody against MCCC2. Recognizes MCCC2 from Human, Mouse. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC; Recommended dilution: WB:1:1000-1:5000, IHC:1:20-1:200

MCCC2 Antibody

DF12655 200ul
EUR 304
Description: MCCC2 Antibody detects endogenous levels of MCCC2.

MCCC2 antibody

70R-18436 50 ul
EUR 435
Description: Rabbit polyclonal MCCC2 antibody

MCCC2 Polyclonal Antibody

28858-100ul 100ul
EUR 252

MCCC2 Polyclonal Antibody

28858-50ul 50ul
EUR 187

MCCC2 Polyclonal Antibody

31405-100ul 100ul
EUR 252

MCCC2 Polyclonal Antibody

31405-50ul 50ul
EUR 187

Mccc2/ Rat Mccc2 ELISA Kit

ELI-48233r 96 Tests
EUR 886

MCCC2 siRNA

20-abx923707
  • EUR 551.00
  • EUR 732.00
  • 15 nmol
  • 30 nmol

MCCC2 siRNA

20-abx923708
  • EUR 551.00
  • EUR 732.00
  • 15 nmol
  • 30 nmol

MCCC2 siRNA

20-abx903187
  • EUR 551.00
  • EUR 732.00
  • 15 nmol
  • 30 nmol

Polyclonal MCCC2 Antibody (Center)

APR10893G 0.1ml
EUR 528
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human MCCC2 (Center). This antibody is tested and proven to work in the following applications:

MCCC2 Polyclonal Conjugated Antibody

C31405 100ul
EUR 397

MCCC2 Polyclonal Conjugated Antibody

C28858 100ul
EUR 397

Polyclonal Goat anti-GST α-form

GST-ANTI-1 50 uL
EUR 280

Polyclonal Goat anti-GST μ-form

GST-ANTI-2 50 uL
EUR 280

Polyclonal Goat anti-GST p-form

GST-ANTI-3 50 uL
EUR 280

MCCC2 Rabbit pAb

A15181-100ul 100 ul
EUR 308

MCCC2 Rabbit pAb

A15181-200ul 200 ul
EUR 459

MCCC2 Rabbit pAb

A15181-20ul 20 ul
EUR 183

MCCC2 Rabbit pAb

A15181-50ul 50 ul
EUR 223

MCCC2 Rabbit pAb

A7990-100ul 100 ul
EUR 308

MCCC2 Rabbit pAb

A7990-200ul 200 ul
EUR 459

MCCC2 Rabbit pAb

A7990-20ul 20 ul
EUR 183

MCCC2 Rabbit pAb

A7990-50ul 50 ul
EUR 223

MCCC2 cloning plasmid

CSB-CL867199HU1-10ug 10ug
EUR 233
Description: A cloning plasmid for the MCCC2 gene.

MCCC2 cloning plasmid

CSB-CL867199HU2-10ug 10ug
EUR 376
Description: A cloning plasmid for the MCCC2 gene.

MCCC2 Blocking Peptide

DF12655-BP 1mg
EUR 195

[One Step] MCCC2 Antibody Kit

RK05665 50 ul
EUR 240

Methylcrotonoyl-CoA Carboxylase 2 (MCCC2) Antibody

abx037012-100ug 100 ug
EUR 391

Methylcrotonoyl-CoA Carboxylase 2 (MCCC2) Antibody

20-abx006595
  • EUR 411.00
  • EUR 592.00
  • EUR 182.00
  • EUR 314.00
  • 100 ul
  • 200 ul
  • 20 ul
  • 50 ul

Methylcrotonoyl-CoA Carboxylase 2 (MCCC2) Antibody

abx030686-400ul 400 ul
EUR 523

Methylcrotonoyl-CoA Carboxylase 2 (MCCC2) Antibody

abx030686-80l 80 µl
EUR 286

Methylcrotonoyl-CoA Carboxylase 2 (MCCC2) Antibody

abx033068-400ul 400 ul
EUR 523

Methylcrotonoyl-CoA Carboxylase 2 (MCCC2) Antibody

abx033068-80l 80 µl
EUR 286

Methylcrotonoyl-CoA Carboxylase 2 (MCCC2) Antibody

20-abx322517
  • EUR 439.00
  • EUR 328.00
  • 100 ul
  • 50 ul

Methylcrotonoyl-CoA Carboxylase 2 (MCCC2) Antibody

abx235048-100ug 100 ug
EUR 509

Mouse MCCC2 shRNA Plasmid

20-abx978856
  • EUR 801.00
  • EUR 1121.00
  • 150 µg
  • 300 µg

Rat MCCC2 shRNA Plasmid

20-abx990287
  • EUR 801.00
  • EUR 1121.00
  • 150 µg
  • 300 µg

Human MCCC2 shRNA Plasmid

20-abx961908
  • EUR 801.00
  • EUR 1121.00
  • 150 µg
  • 300 µg

Mouse Mccc2 ELISA KIT

ELI-46423m 96 Tests
EUR 865

MCCC2 ELISA KIT|Human

EF000692 96 Tests
EUR 689

Human MCCC2 ELISA KIT

ELI-15862h 96 Tests
EUR 824

MCCC2 Recombinant Protein (Human)

RP018958 100 ug Ask for price

MCCC2 Recombinant Protein (Human)

RP018961 100 ug Ask for price

MCCC2 Recombinant Protein (Mouse)

RP149789 100 ug Ask for price

MCCC2 Recombinant Protein (Rat)

RP211097 100 ug Ask for price

Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) Antibody

20-abx131298
  • EUR 439.00
  • EUR 133.00
  • EUR 1233.00
  • EUR 592.00
  • EUR 328.00
  • 100 ug
  • 10 ug
  • 1 mg
  • 200 ug
  • 50 ug

Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) Antibody

20-abx131299
  • EUR 453.00
  • EUR 133.00
  • EUR 1302.00
  • EUR 620.00
  • EUR 342.00
  • 100 ug
  • 10 ug
  • 1 mg
  • 200 ug
  • 50 ug

Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) Antibody

20-abx113712
  • EUR 732.00
  • EUR 398.00
  • 150 ul
  • 50 ul

MCCC2 ELISA Kit (Human) (OKCD00916)

OKCD00916 96 Wells
EUR 831
Description: Description of target: Carboxyltransferase subunit of the 3-methylcrotonyl-CoA carboxylase, an enzyme that catalyzes the conversion of 3-methylcrotonyl-CoA to 3-methylglutaconyl-CoA, a critical step for leucine and isovaleric acid catabolism.1 Publication <p>Manually curated information for which there is published experimental evidence.</p> <p><a href="/manual/evidences#ECO:0000269">More…</a></p> Manual assertion based on experiment iniRef.9"Expression, purification, characterization of human 3-methylcrotonyl-CoA carboxylase (MCCC)."_x005F_x005F_x000D_Chu C.H., Cheng D._x005F_x005F_x000D_Protein Expr. Purif. 53:421-427(2007) [PubMed] [Europe PMC] [Abstract]Cited for: FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES. ;Species reactivity: Human;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich Immunoassay;Sensitivity: < 0.141 ng/mL

Mccc2 ORF Vector (Mouse) (pORF)

ORF049931 1.0 ug DNA
EUR 506

Mccc2 ORF Vector (Rat) (pORF)

ORF070367 1.0 ug DNA
EUR 506

MCCC2 ORF Vector (Human) (pORF)

ORF006320 1.0 ug DNA
EUR 95

MCCC2 ORF Vector (Human) (pORF)

ORF006321 1.0 ug DNA
EUR 95

Mccc2 sgRNA CRISPR Lentivector set (Mouse)

K3464301 3 x 1.0 ug
EUR 339

MCCC2 sgRNA CRISPR Lentivector set (Human)

K1278301 3 x 1.0 ug
EUR 339

Mccc2 sgRNA CRISPR Lentivector set (Rat)

K7573601 3 x 1.0 ug
EUR 339

Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) Polyclonal Antibody (Human)

4-PAD690Hu01
  • EUR 261.00
  • EUR 2734.00
  • EUR 676.00
  • EUR 330.00
  • EUR 220.00
  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Human Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2)

Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2) Polyclonal Antibody (Mouse)

4-PAD690Mu01
  • EUR 251.00
  • EUR 2576.00
  • EUR 640.00
  • EUR 316.00
  • EUR 215.00
  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Mouse Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2)

Recombinant Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2)

4-RPD690Hu01
  • EUR 526.50
  • EUR 244.00
  • EUR 1699.36
  • EUR 633.12
  • EUR 1166.24
  • EUR 415.00
  • EUR 4098.40
  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Human Methylcrotonoyl Coenzyme A Carboxylase 2 expressed in: E.coli

Recombinant Methylcrotonoyl Coenzyme A Carboxylase 2 (MCCC2)

4-RPD690Mu01
  • EUR 544.42
  • EUR 248.00
  • EUR 1766.56
  • EUR 655.52
  • EUR 1211.04
  • EUR 427.00
  • EUR 4266.40
  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Mouse Methylcrotonoyl Coenzyme A Carboxylase 2 expressed in: E.coli

Mccc2 sgRNA CRISPR Lentivector (Mouse) (Target 1)

K3464302 1.0 ug DNA
EUR 154

Mccc2 sgRNA CRISPR Lentivector (Mouse) (Target 2)

K3464303 1.0 ug DNA
EUR 154

Mccc2 sgRNA CRISPR Lentivector (Mouse) (Target 3)

K3464304 1.0 ug DNA
EUR 154

MCCC2 sgRNA CRISPR Lentivector (Human) (Target 1)

K1278302 1.0 ug DNA
EUR 154

MCCC2 sgRNA CRISPR Lentivector (Human) (Target 2)

K1278303 1.0 ug DNA
EUR 154

MCCC2 sgRNA CRISPR Lentivector (Human) (Target 3)

K1278304 1.0 ug DNA
EUR 154

Mccc2 sgRNA CRISPR Lentivector (Rat) (Target 1)

K7573602 1.0 ug DNA
EUR 154

Mccc2 sgRNA CRISPR Lentivector (Rat) (Target 2)

K7573603 1.0 ug DNA
EUR 154

Mccc2 sgRNA CRISPR Lentivector (Rat) (Target 3)

K7573604 1.0 ug DNA
EUR 154

Mccc2 3'UTR GFP Stable Cell Line

TU262953 1.0 ml Ask for price

Mccc2 3'UTR GFP Stable Cell Line

TU162987 1.0 ml Ask for price

Mccc2 3'UTR Luciferase Stable Cell Line

TU212953 1.0 ml Ask for price

MCCC2 3'UTR GFP Stable Cell Line

TU063103 1.0 ml
EUR 1521

Mccc2 3'UTR Luciferase Stable Cell Line

TU112987 1.0 ml Ask for price

MCCC2 3'UTR Luciferase Stable Cell Line

TU013103 1.0 ml
EUR 1521

MCCC2 Protein Vector (Human) (pPB-C-His)

PV025277 500 ng
EUR 329

MCCC2 Protein Vector (Human) (pPB-N-His)

PV025278 500 ng
EUR 329

MCCC2 Protein Vector (Human) (pPM-C-HA)

PV025279 500 ng
EUR 329

MCCC2 Protein Vector (Human) (pPM-C-His)

PV025280 500 ng
EUR 329

MCCC2 Protein Vector (Human) (pPB-C-His)

PV025281 500 ng
EUR 329

MCCC2 Protein Vector (Human) (pPB-N-His)

PV025282 500 ng
EUR 329

MCCC2 Protein Vector (Human) (pPM-C-HA)

PV025283 500 ng
EUR 329

MCCC2 Protein Vector (Human) (pPM-C-His)

PV025284 500 ng
EUR 329

MCCC2 Protein Vector (Rat) (pPB-C-His)

PV281466 500 ng
EUR 603

MCCC2 Protein Vector (Rat) (pPB-N-His)

PV281467 500 ng
EUR 603

MCCC2 Protein Vector (Rat) (pPM-C-HA)

PV281468 500 ng
EUR 603

MCCC2 Protein Vector (Rat) (pPM-C-His)

PV281469 500 ng
EUR 603

The tactic provides the benefit of being carried out on stay cells, assuaging the necessity for a recombinant supply of the receptor. Moreover, time-resolved measurements, along with permitting the willpower of the affinity of the studied drug to its goal, give entry to the binding kinetics thereby offering a full characterization of the system. On this research, we additionally in contrast time-resolved LigandTracer information with end-point OkayD willpower from circulation cytometry experiments and hypothesize that discrepancies between these two approaches, once they exist, typically come from circulation cytometry titration curves being acquired previous to full equilibration of the system.

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