Development of flow cytometry assays for measuring cell-membrane enzyme activity on individual cells.

Development of flow cytometry assays for measuring cell-membrane enzyme activity on individual cells.

Background: Cell membranes expressing enzymes such as ADAM (disintegrin and metalloproteinases) superfamily members are considered key catalyst vital cellular functions. To directly measure this enzyme and determine their relationship with certain cells and function, individual-cell membrane-bound enzyme activity tests are needed, but not available.

Methods: We developed two such tests, using fluorescence resonance energy transfer (FRET) peptide substrate (FPS) and flow cytometry. One test measured the natural processing of live-cell fluorescent products FPS and its binding to membrane-bound enzyme-cell individuals. Test measured other treatment of special-bound generation and glutaraldehyde cross FPS, and consequent fluorescent products are coupled to a membrane-bound enzyme-cell individuals.

Results: confocal microscopy-imaging shows that proteolytic processing of FPS selective and labeled occur in the cell membrane of individual cells. New test tool measured increase FPS specifically related fluorescent products in the cell-substrate concentration-, courtesy of temperature and time dependent. Most of the products processed remain relevant FPS fluorescent cells after washing the cells, suggesting they bind to cells expressing membrane enzymes. Assays measure the level higher than the FPS neon related products on the wild-type cells of ADAM10 knockout mouse fibroblasts and in human monocytes than lymphocytes, which correlated with the presence and expression levels of ADAM10 in the cell membrane, respectively.

Furthermore, test enzyme activity can be combined with anti-ADAM10 fluorescent antibody staining to co-label and more direct association enzyme activity and the level of ADAM10 protein in the cell membrane of individual cells. Conclusion: We report two new test for measuring cell membrane-anchored enzyme activity in individual cells, and the potential for its use to directly study the biology of specific protease-expressing cell surface.

Development of flow cytometry assays for measuring cell-membrane enzyme activity on individual cells.
Development of flow cytometry assays for measuring cell-membrane enzyme activity on individual cells.

Building a model system to evaluate CAR T cell therapy using dogs with spontaneous diffuse large B-cell lymphoma.

Some rodent and primate preclinical studies have CAR T cells advanced into the clinic. However, there is no single model to accurately reflect the challenge CAR T effective therapy in human cancer patients. To evaluate the effectiveness of CAR T cell next generation that aims to overcome barriers to long-lasting removal of the tumor, we developed a system to evaluate the CAR T cells in dogs with spontaneous cancers.

Here we report the results of this system and a pilot experiment using CAR T cells to treat dogs spread large B-cell lymphoma (DLBCL). We designed and produced CD20-targeting, a second-generation dog CAR T cells to functional evaluation in vitro and in vivo using lentivectors parallel to the human T cell manufacturing CAR. A trial is the first-in-five species of dogs with DLBCL treated with CAR T do.

Canine CAR T cell antigen-specific function and kill CD20 + targets. Circulation CAR T cells were detected post-infusion, however, the induction of anti-mouse antibody dog ​​(CAMA) is associated with a loss of CAR T cells specific selection pressures in tumors were observed following CD20 + CAR T cell therapy, culminating in a runaway antigen and the emergence of CD20-disease. the survival time of patients correlated with the ex vivo expansion of the product.

GOAT ANTI MOUSE IgG

MBS235208-5x1mg 5x1mg
EUR 925

Goat anti Mouse IgG

MBS539273-20mg 20mg
EUR 440

Goat anti Mouse IgG

MBS539273-5x20mg 5x20mg
EUR 1820

Goat Anti Mouse IgG

MBS8503086-01mg 0.1mg
EUR 305

Goat Anti Mouse IgG

MBS8503086-5x01mg 5x0.1mg
EUR 1220

Goat anti Mouse IgG (Fc)

MBS315381-2mg 2mg
EUR 315

Goat anti Mouse IgG (Fc)

MBS315381-5x2mg 5x2mg
EUR 1240

Goat anti Mouse IgG (Fc)

MBS316344-5mg 5mg
EUR 440

Goat anti Mouse IgG (Fc)

MBS316344-5x5mg 5x5mg
EUR 1800

Goat anti Mouse IgG (Fc)

MBS316504-2mL 2mL
EUR 590

Goat anti Mouse IgG (Fc)

MBS316504-5x2mL 5x2mL
EUR 2485

Goat anti Mouse IgG (Fc)

MBS316505-2mg 2mg
EUR 505

Goat anti Mouse IgG (Fc)

MBS316505-5x2mg 5x2mg
EUR 2100

Goat anti Mouse IgG (Fc)

MBS316557-1mg 1mg
EUR 490

Goat anti Mouse IgG (Fc)

MBS316557-5x1mg 5x1mg
EUR 2030

GOAT ANTI MOUSE IgG (Fc)

MBS235248-1mg 1mg
EUR 265

GOAT ANTI MOUSE IgG (Fc)

MBS235248-5x1mg 5x1mg
EUR 1020

Goat anti Mouse IgG (HRP)

43C-CB1569 1 mg
EUR 181
Description: Goat anti Mouse IgG secondary antibody  (HRP)

Goat anti Mouse IgG + IgM

41R-1063 1 mg
EUR 115
Description: Goat anti Mouse IgG + IgM secondary antibody

Goat anti Mouse IgG + IgM

41R-1064 1 mg
EUR 176
Description: Goat anti Mouse IgG + IgM secondary antibody

Goat anti Mouse IgG Fab

MBS571371-1mL 1mL
EUR 260

Goat anti Mouse IgG Fab

MBS571371-5x1mL 5x1mL
EUR 1015

Goat anti Mouse IgG Fab

MBS571162-10mg 10mg
EUR 625

Goat anti Mouse IgG Fab

MBS571162-5x10mg 5x10mg
EUR 2665

Goat anti Mouse IgG (HRP)

MBS539860-1mg 1mg
EUR 300

Goat anti Mouse IgG (HRP)

MBS539860-5x1mg 5x1mg
EUR 1200

Goat anti Mouse IgG (FITC)

43R-1359 2 mg
EUR 167
Description: Goat anti Mouse IgG secondary antibody  (FITC)

Goat Anti Mouse IgG: FITC

MBS225563-1mg 1mg
EUR 265

Goat Anti Mouse IgG: FITC

MBS225563-5x1mg 5x1mg
EUR 1020

Goat Anti Mouse IgG-HRP

E61I00503 1mg
EUR 345

Goat anti Mouse IgG (gamma)

MBS315382-2mg 2mg
EUR 290

Goat anti Mouse IgG (gamma)

MBS315382-5x2mg 5x2mg
EUR 1135

Goat Anti Mouse IgG-HRP

MBS8502251-1mg 1mg
EUR 425

Goat Anti Mouse IgG-HRP

MBS8502251-5x1mg 5x1mg
EUR 1755

Goat Anti Mouse IgG -Biotin

E61I00502 1mg
EUR 345

Goat Anti Mouse IgG-FITC

E61I00504 1mg
EUR 431.25

Goat Anti Mouse IgG-FITC

MBS8503746-1mg 1mg
EUR 510

Goat Anti Mouse IgG-FITC

MBS8503746-5x1mg 5x1mg
EUR 2140

Goat Anti Mouse IgG-Biotin

MBS8503038-5x1mg 5x1mg
EUR 1755

Goat Anti Mouse IgG-Biotin

MBS8503038-1mg 1mg
EUR 425

Goat anti Mouse IgG (rhodamine)

43R-1360 1 mg
EUR 116
Description: Goat anti Mouse IgG secondary antibody  (rhodamine)

Goat anti Mouse IgG (H + L)

41C-CB1501 2 mg
EUR 193
Description: Goat anti Mouse IgG (H + L) secondary antibody

Goat anti Mouse IgG (H + L)

41R-1051 2 mg
EUR 127
Description: Goat anti Mouse IgG (H + L) secondary antibody

Goat anti Mouse IgG (H + L)

41R-1052 2 mg
EUR 115
Description: Goat anti Mouse IgG (H + L) secondary antibody

Goat anti Mouse IgG (H + L)

41R-1053 2 mg
EUR 128
Description: Goat anti Mouse IgG (H + L) secondary antibody

Goat anti Mouse IgG (H + L)

41R-1054 2 mg
EUR 176
Description: Goat anti Mouse IgG (H + L) secondary antibody

Goat anti Mouse IgG (H + L)

41R-1055 2 mg
EUR 299
Description: Goat anti Mouse IgG (H + L) secondary antibody

Goat anti Mouse IgG (H + L)

41R-1056 1 mg
EUR 133
Description: Goat anti Mouse IgG (H + L) secondary antibody

Goat anti Mouse IgG (H + L)

41R-1057 1 mg
EUR 167
Description: Goat anti Mouse IgG (H + L) secondary antibody

Goat anti Mouse IgG (H + L)

41R-1100 2 mg
EUR 176
Description: Goat anti Mouse IgG (H + L) secondary antibody

Goat anti Mouse IgG (H + L)

40-GM25 100 ml
EUR 399
Description: Goat anti Mouse IgG secondary antibody (H + L)

Goat anti- mouse IgG(H+L)

FNab09848 100µg
EUR 658.5
Description: Antibody raised against Goat mouse IgG(H+L)

Goat anti Mouse IgG (H&L)

MBS315376-2mg 2mg
EUR 265

Goat anti Mouse IgG (H&L)

MBS315376-5x2mg 5x2mg
EUR 1020

Goat anti Mouse IgG (H&L)

MBS315378-2mg 2mg
EUR 265

Goat anti Mouse IgG (H&L)

MBS315378-5x2mg 5x2mg
EUR 1020

Goat anti Mouse IgG (H&L)

MBS315379-1mg 1mg
EUR 265

Goat anti Mouse IgG (H&L)

MBS315379-5x1mg 5x1mg
EUR 1020

Goat anti Mouse IgG (H&L)

MBS316345-5mg 5mg
EUR 280

Goat anti Mouse IgG (H&L)

MBS316345-5x5mg 5x5mg
EUR 1090

Goat anti Mouse IgG (H&L)

MBS316442-15mg 1.5mg
EUR 470

Goat anti Mouse IgG (H&L)

MBS316442-5x15mg 5x1.5mg
EUR 1935

Goat anti Mouse IgG (H&L)

MBS316751-1mg 1mg
EUR 310

Goat anti Mouse IgG (H&L)

MBS316751-5x1mg 5x1mg
EUR 1230

GOAT ANTI MOUSE IGG H&L

MBS316824-5mg 5mg
EUR 440

GOAT ANTI MOUSE IGG H&L

MBS316824-5x5mg 5x5mg
EUR 1800

GOAT ANTI MOUSE IgG (H/L)

MBS235231-2mg 2mg
EUR 440

GOAT ANTI MOUSE IgG (H/L)

MBS235231-5x2mg 5x2mg
EUR 1815

Goat anti Mouse IgG (H + L)

MBS539137-100mL 100mL
EUR 870

Goat anti Mouse IgG (H + L)

MBS539137-5x100mL 5x100mL
EUR 3760

Goat anti Mouse IgG (H + L)

MBS539856-2mg 2mg
EUR 370

Goat anti Mouse IgG (H + L)

MBS539856-5x2mg 5x2mg
EUR 1525

Goat anti Mouse IgG (Fab'2)

41C-CJ0207 1 mg
EUR 279
Description: Goat anti Mouse IgG secondary antibody (Fab'2)

Goat anti Mouse IgG (Fab'2)

41C-CJ0213 1 mg
EUR 333
Description: Goat anti Mouse IgG secondary antibody (Fab'2)

GOAT ANTI MOUSE IgG (Fc):HRP

MBS235188-1mg 1mg
EUR 320

GOAT ANTI MOUSE IgG (Fc):HRP

MBS235188-5x1mg 5x1mg
EUR 1255

Goat anti Mouse IgG (Fab'2)

MBS539801-1mg 1mg
EUR 700

Goat anti Mouse IgG (Fab'2)

MBS539801-5x1mg 5x1mg
EUR 2990

Goat anti Mouse IgG (Fab'2)

MBS536022-1mg 1mg
EUR 575

Goat anti Mouse IgG (Fab'2)

MBS536022-5x1mg 5x1mg
EUR 2435

Goat anti Mouse IgG + IgM (HRP)

43R-IG051hrp 1 mg
EUR 181
Description: Goat anti Mouse IgG + IgM secondary antibody (HRP)

GOAT ANTI MOUSE IgG (Fc):FITC

MBS235143-1mg 1mg
EUR 290

GOAT ANTI MOUSE IgG (Fc):FITC

MBS235143-5x1mg 5x1mg
EUR 1135

Goat anti Mouse IgG + IgM (HRP)

MBS539229-1mg 1mg
EUR 300

Goat anti Mouse IgG + IgM (HRP)

MBS539229-5x1mg 5x1mg
EUR 1200

Goat anti Mouse IgG (Alk Phos)

43R-1357 1 mg
EUR 208
Description: Goat anti Mouse IgG secondary antibody  (Alk Phos)

Goat anti Mouse IgG + IgM (FITC)

43R-1388 1 mg
EUR 167
Description: Goat anti Mouse IgG + IgM secondary antibody (FITC)

Goat anti Mouse IgG Fc (biotin)

43R-1358 2 mg
EUR 208
Description: Goat anti Mouse IgG secondary antibody (biotin)

GOAT ANTI MOUSE IGG, INTACT IGG, HRP, 2 ML

9450714 1ML
EUR 207.65

GOAT ANTI MOUSE IGG, INTACT IGG, FITC, 2 MG

9450713 1MG
EUR 169.58

GOAT ANTI MOUSE IgG (RAT ADSORBED)

MBS235255-05mg 0.5mg
EUR 445

GOAT ANTI MOUSE IgG (RAT ADSORBED)

MBS235255-5x05mg 5x0.5mg
EUR 1830

Goat anti Mouse IgG + IgM (rhodamine)

43R-1389 1 mg
EUR 167
Description: Goat anti Mouse IgG + IgM secondary antibody (rhodamine)

GOAT ANTI MOUSE IGG, INTACT IGG, UNCONJ., 2 MG

9450710 1MG
EUR 153.48

GOAT ANTI MOUSE IGG, INTACT IGG, BIOTIN, 2 ML

9450712 1ML
EUR 207.65

IgG Antibody (Goat anti Mouse) (Fc)

GWB-B6FACE 2 ml Ask for price

IgG Antibody (Goat anti Mouse) (Fc)

GWB-CD3640 2 ml Ask for price

IgG Antibody (Goat anti Mouse) (Fc)

GWB-BEA930 1 mg Ask for price

IgG Antibody (Goat anti Mouse) (Fc)

GWB-D17EA6 2 mg Ask for price

IgG Antibody (Goat anti Mouse) (Fc)

GWB-D6570C 1 mg Ask for price

IgG Antibody (Goat anti Mouse) (Fc)

GWB-D8C9F1 2 mg Ask for price

IgG Antibody (Goat anti Mouse) (Fc)

GWB-49F3C9 1 ml Ask for price

IgG Antibody (Goat anti Mouse) (Fc)

GWB-1C6569 5 mg Ask for price

IgG Antibody (Fab) (Goat Anti Mouse)

GWB-5815F1 2 mg Ask for price

Goat Anti-Rabbit & Mouse IgG-HRP

MBS435024-01mL 0.1mL
EUR 180

Goat Anti-Rabbit & Mouse IgG-HRP

MBS435024-1mL 1mL
EUR 300

Goat Anti-Rabbit & Mouse IgG-HRP

MBS435024-5x1mL 5x1mL
EUR 1020

GOAT ANTI MOUSE IgG (MULTISPECIES ADSORBED)

MBS235223-1mg 1mg
EUR 460

GOAT ANTI MOUSE IgG (MULTISPECIES ADSORBED)

MBS235223-5x1mg 5x1mg
EUR 1890

Goat Anti Mouse IgG, F(ab’)2

FNSA-0030 500 uL
EUR 367.92
Description: Goat Anti Mouse IgG, F(ab’)2 secondary antibody

GOAT ANTI RAT IgG (MOUSE ADSORBED)

MBS235254-05mg 0.5mg
EUR 290

GOAT ANTI RAT IgG (MOUSE ADSORBED)

MBS235254-5x05mg 5x0.5mg
EUR 1135

Goat anti Mouse IgG (H + L) (HRP)

43-GM30 2 mg
EUR 264
Description: Goat anti Mouse IgG (H + L) secondary antibody (HRP)

Goat anti Mouse IgG (Fab'2) (PE)

43C-CJ0212 500 µg
EUR 551
Description: Goat anti Mouse IgG secondary antibody (Fab'2) (PE)

Goat anti Mouse IgG (Fab'2) (PE)

43C-CJ0218 500 µg
EUR 551
Description: Goat anti Mouse IgG secondary antibody (Fab'2) (PE)

Goat anti Mouse IgG (H + L) (HRP)

43R-1318 1 mg
EUR 117
Description: Goat anti Mouse IgG (H + L) secondary antibody (HRP)

Goat anti Mouse IgG (H + L) (HRP)

43R-1324 1 mg
EUR 116
Description: Goat anti Mouse IgG (H + L) secondary antibody (HRP)

Goat anti Mouse IgG (H + L) (HRP)

43R-1330 1 mg
EUR 128
Description: Goat anti Mouse IgG (H + L) secondary antibody (HRP)

Goat anti Mouse IgG (H + L) (HRP)

43R-1336 1 mg
EUR 190
Description: Goat anti Mouse IgG (H + L) secondary antibody (HRP)

Goat anti Mouse IgG (H + L) (HRP)

43R-1342 1 mg
EUR 336
Description: Goat anti Mouse IgG (H + L) secondary antibody (HRP)

Goat anti Mouse IgG (H + L) (HRP)

43R-1348 500 ug
EUR 151
Description: Goat anti Mouse IgG (H + L) secondary antibody (HRP)

Goat anti Mouse IgG (H + L) (HRP)

43R-1354 500 ug
EUR 190
Description: Goat anti Mouse IgG (H + L) secondary antibody (HRP)

Goat anti Mouse IgG (H + L) (HRP)

43R-1610 1 mg
EUR 192
Description: Goat anti Mouse IgG (H + L) secondary antibody (HRP)

Goat anti Mouse IgG + IgM (H + L)

40C-CB1569 1 mg
EUR 196
Description: Goat anti Mouse IgG + IgM (H + L) secondary antibody

GOAT ANTI MOUSE IgG (H/L):HRP

MBS235171-1mg 1mg
EUR 300

GOAT ANTI MOUSE IgG (H/L):HRP

MBS235171-5x1mg 5x1mg
EUR 1175

GOAT ANTI MOUSE IgG (H/L):HRP

MBS235172-2mg 2mg
EUR 360

GOAT ANTI MOUSE IgG (H/L):HRP

MBS235172-5x2mg 5x2mg
EUR 1445

GOAT ANTI MOUSE IgG (H/L):HRP

MBS219959-2mL 2mL
EUR 645

GOAT ANTI MOUSE IgG (H/L):HRP

MBS219959-5x2mL 5x2mL
EUR 2720

Goat anti Mouse IgG (Fab'2) (PE)

MBS538836-05mg 0.5mg
EUR 1025

Goat anti Mouse IgG (Fab'2) (PE)

MBS538836-5x05mg 5x0.5mg
EUR 4460

Goat anti Mouse IgG (Fab'2) (PE)

MBS538841-05mg 0.5mg
EUR 1025

Goat anti Mouse IgG (Fab'2) (PE)

MBS538841-5x05mg 5x0.5mg
EUR 4460

Goat anti Mouse IgG (H + L) (HRP)

MBS539266-2mg 2mg
EUR 500

Goat anti Mouse IgG (H + L) (HRP)

MBS539266-5x2mg 5x2mg
EUR 2100

Goat anti Mouse IgG (Fab'2) (HRP)

43C-CJ0211 500 µg
EUR 551
Description: Goat anti Mouse IgG secondary antibody (Fab'2) (HRP)

Goat anti Mouse IgG (Fab'2) (HRP)

43C-CJ0217 500 µg
EUR 44
Description: Goat anti Mouse IgG secondary antibody (Fab'2) (HRP)

Goat anti Mouse IgG (H + L) (FITC)

43R-1317 2 mg
EUR 123
Description: Goat anti Mouse IgG (H + L) secondary antibody (FITC)

Goat anti Mouse IgG (H + L) (FITC)

43R-1323 2 mg
EUR 167
Description: Goat anti Mouse IgG (H + L) secondary antibody (FITC)

Goat anti Mouse IgG (H + L) (FITC)

43R-1329 2 mg
EUR 189
Description: Goat anti Mouse IgG (H + L) secondary antibody (FITC)

Goat anti Mouse IgG (H + L) (FITC)

43R-1335 1 mg
EUR 167
Description: Goat anti Mouse IgG (H + L) secondary antibody (FITC)

Goat anti Mouse IgG (H + L) (FITC)

43R-1347 500 ug
EUR 133
Description: Goat anti Mouse IgG (H + L) secondary antibody (FITC)

Goat anti Mouse IgG (H + L) (FITC)

43R-1353 500 ug
EUR 167
Description: Goat anti Mouse IgG (H + L) secondary antibody (FITC)

Goat anti Mouse IgG (H + L) (FITC)

43R-1611 1 mg
EUR 192
Description: Goat anti Mouse IgG (H + L) secondary antibody (FITC)

Changing the manufactured products increased efficiency transduction and tilted toward such memory T cell phenotype CAR dogs. CAR T cell manufacturing dogs lentivector functional use is feasible. CAR effectively challenge comparable to existing T cell therapy, demonstrating its relevance in the future to inform the design of human clinical trials.

Nathaniel

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