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Current position:Product Center > Cell lines > TNF > TL1A:DR3(TNFRSF25) > Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line
Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line
Description
Cat. No: GM-C30291
Product: Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line


Tumor necrosis factor-like ligand 1A (TL1A), or TNFSF15, is a cytokine primarily expressed by endothelial cells. In T cells, it functions as a co-stimulator, boosting IL-2 reactivity and pro-inflammatory cytokine secretion. TL1A is the sole ligand for death receptor 3 (DR3 or TNFRSF25), a TNF receptor family member that induces apoptosis upon T cell activation. Blocking the TL1A-DR3 interaction is a potential target for chronic immune disease therapies. Furthermore, DR3 agonistic antibodies can reduce regulatory T cell suppression and enhance CD4+ T cell activity in mouse melanoma models, indicating their potential as treatments for solid tumors.
Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line is a clonal stable Jurkat cell line constructed using lentiviral technology, constitutively expressing the Mouse TNFRSF25(DR3), along with signal-dependent expression of a luciferase reporter gene.
It has two application scenarios:
First, by adding a DR3 agonist drug to activate the downstream signaling pathways in reporter gene cells, and measuring the fluorescence signal to determine the expression of luciferase, it can be used to screen or validate agonist drugs targeting mouse DR3.
Second, by adding a TL1A antagonist antibody to block the downstream mouse DR3 signaling activated by TL1A, and measuring the fluorescence signal to determine the expression of luciferase, it can be used to screen or validate antagonist drugs targeting TL1A.
Data
Response to Anti-TNFRSF25(DR3) hIgG1 Antibody(PTX-35). The Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line (Cat. GM-C30291) at a concentration of 1E5 cells/well (96-well format) was stimulated with serial dilutions of Anti-TNFRSF25(DR3) hIgG1 Antibody(PTX-35) (Cat. GM-58913AB) in assay buffer (RPMI 1640+1% FBS+1% P.S) for 7 hours. The firefly luciferase activity was measured using the GMOne-Step Luciferase Reporter Gene Assay Kit (Cat. GM-040503). The maximum induction fold was approximately [3.4]. Data are shown by drug mass concentration.
Response to Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605). Serial dilutions of the Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605) (Cat. GM-59479AB) were incubated with 1E4 cells/well of the Mouse_TNFSF15(TL1A) HEK-293 Cell Line (Cat. GM-C31356) in a 96-well plate for 1 hour in assay buffer (RPMI 1640 + 1% FBS + 1% P.S). Subsequently, the Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line (Cat. GM-C30291) at a concentration of 1E5 cells/well was added, and the co-culture proceeded for an additional 7 hours. Firefly luciferase activity is then measured using the GMOne-Step Luciferase Reporter Gene Assay Kit (Cat. GM-040503). The results indicated a maximum blocking fold of approximately [172.2]. Data are shown by drug mass concentration.
Response to Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605). Serial dilutions of the Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605) (Cat. GM-59479AB) were incubated with 2E3 cells/well of the H_TNFSF15(TL1A) CHO-K1 Cell Line (Cat. GM-C19170) in a 96-well plate for 1 hour in assay buffer (RPMI 1640 + 1% FBS + 1% P.S). Subsequently, the Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line (Cat. GM-C30291) at a concentration of 1E5 cells/well was added, and the co-culture proceeded for an additional 6 hours. Firefly luciferase activity is then measured using the GMOne-Step Luciferase Reporter Gene Assay Kit (Cat. GM-040503). The results indicated a maximum blocking fold of approximately [26.0]. Data are shown by drug mass concentration.
Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line (Cat. GM-C30291) was determined by flow cytometry using Anti-TNFRSF25(DR3) hIgG1 Antibody(PTX-35) (Cat. GM-58913AB).
Specifications
Cat. NoGM-C30291
ProductMouse_TNFRSF25(DR3) Reporter Jurkat Cell Line
Product Format1 vial of frozen cells
Quantity5E6 Cells per vial,1 mL
Storage ConditionsLiquid nitrogen immediately upon receipt
Recovery MediumRPMI 1640+10% FBS+1% P.S
Growth mediumRPMI 1640+10% FBS+1% P.S+3.5 μg/mL Blasticidin+0.75 μg/mL Puromycin
NoteNone
Freezing Medium90% FBS+10% DMSO
Growth propertiesSuspension
Growth Conditions37°C, 5% CO₂
Safety considerationsBiosafety Level 2
Mycoplasma TestingThe cell line has been screened to confirm the absence of Mycoplasma species.
Materials
ReagentOrdering Information
BlasticidinGenomeditech/GM-040404
PuromycinGenomeditech/GM-040401
Pen/StrepThermo/15140-122
Fetal Bovine SerumCegrogen biotech/A0500-3010
RPMI 1640VivaCell/C3010-0500
Clear Flat-Bottom Immuno Nonsterile 96-Well PlatesThermo/442404
H_TNFSF15(TL1A) CHO-K1 Cell LineGenomeditech/GM-C19170
Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605)Genomeditech/GM-59479AB
Anti-TNFRSF25(DR3) hIgG1 Antibody(PTX-35)Genomeditech/GM-58913AB
GMOne-Step Luciferase Reporter Gene Assay KitGenomeditech/GM-040503
Cell Culture

Cell Recovery

Recovery Medium: RPMI 1640+10% FBS+1% P.S

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.

a)       Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 - 3 minutes).

b)       Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.

c)       Transfer the vial contents to a centrifuge tube containing 5.0 mL complete culture medium. And spin at approximately 176 x g for 5 minutes. Discard supernatant.

d)       Resuspend cell pellet with the recommended complete medium. And dispense the suspension into 1 - 2 T-25 culture flasks.

e)       Incubate the culture at 37°C in a suitable incubator. A 5% CO₂ in air atmosphere is recommended if using the medium described on this product sheet.

Cell Freezing

Freezing Medium: 90% FBS+10% DMSO

a)          Centrifuge at 176 x g for 3 minutes to collect cells.

b)         Resuspend the cells in pre-cooled freezing medium and adjust the cell density to 5E6 cells/mL.

c)          Aliquot 1 mL into each vial.

d)         Place the vial in a controlled-rate freezing container and store at -80°C for at least 1 day, then transfer to liquid nitrogen as soon as possible.

Cell passage

Growth medium: RPMI 1640+10% FBS+1% P.S+3.5 μg/mL Blasticidin+0.75 μg/mL Puromycin

Approximately 48-72 hours after the initial thawing, the cells can be passaged for the first time. After this initial passage, the culture medium can be adjusted to growth medium supplemented with antibiotics. If cells are not passaged within 48 hours, it is recommended to add some fresh recovery medium and place the flask horizontally.

a)          When the cell density reaches 1.5 - 2E6 cells/mL, subculture the cells. Do not allow the cell density to exceed 2E6 cells/mL.

b)         It is recommended to use T-25 flasks for subculturing.

c)          These cells are suspension cells, and it is recommended to use the "half-medium change" method to maintain optimal cell conditions during passaging.

d)         During passaging, you can directly add fresh growth medium to the culture flask, gently pipette to resuspend the cells, and then transfer the cell suspension to a new T-25 flask for continued culture.

Subcultivation Ratio: Maintain cultures at a cell concentraion between 3E5 and 1E6 viable cells/mL.

Medium Renewal: Every 2 to 3 days

Notes

a)          These cells are sensitive to density, so please ensure that the cell density is maintained within an appropriate range during culture and subculturing.

b)         During the first passage, pay attention to the nutrient supply; if not subculturing, make sure to add fresh recovery medium every other day as needed.

 

Current Position:Product Center > Cell lines > TNF > TL1A:DR3(TNFRSF25) > Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line
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Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line
Description
Cat. No: GM-C30291
Product: Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line


Tumor necrosis factor-like ligand 1A (TL1A), or TNFSF15, is a cytokine primarily expressed by endothelial cells. In T cells, it functions as a co-stimulator, boosting IL-2 reactivity and pro-inflammatory cytokine secretion. TL1A is the sole ligand for death receptor 3 (DR3 or TNFRSF25), a TNF receptor family member that induces apoptosis upon T cell activation. Blocking the TL1A-DR3 interaction is a potential target for chronic immune disease therapies. Furthermore, DR3 agonistic antibodies can reduce regulatory T cell suppression and enhance CD4+ T cell activity in mouse melanoma models, indicating their potential as treatments for solid tumors.
Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line is a clonal stable Jurkat cell line constructed using lentiviral technology, constitutively expressing the Mouse TNFRSF25(DR3), along with signal-dependent expression of a luciferase reporter gene.
It has two application scenarios:
First, by adding a DR3 agonist drug to activate the downstream signaling pathways in reporter gene cells, and measuring the fluorescence signal to determine the expression of luciferase, it can be used to screen or validate agonist drugs targeting mouse DR3.
Second, by adding a TL1A antagonist antibody to block the downstream mouse DR3 signaling activated by TL1A, and measuring the fluorescence signal to determine the expression of luciferase, it can be used to screen or validate antagonist drugs targeting TL1A.
Data
Response to Anti-TNFRSF25(DR3) hIgG1 Antibody(PTX-35). The Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line (Cat. GM-C30291) at a concentration of 1E5 cells/well (96-well format) was stimulated with serial dilutions of Anti-TNFRSF25(DR3) hIgG1 Antibody(PTX-35) (Cat. GM-58913AB) in assay buffer (RPMI 1640+1% FBS+1% P.S) for 7 hours. The firefly luciferase activity was measured using the GMOne-Step Luciferase Reporter Gene Assay Kit (Cat. GM-040503). The maximum induction fold was approximately [3.4]. Data are shown by drug mass concentration.
Response to Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605). Serial dilutions of the Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605) (Cat. GM-59479AB) were incubated with 1E4 cells/well of the Mouse_TNFSF15(TL1A) HEK-293 Cell Line (Cat. GM-C31356) in a 96-well plate for 1 hour in assay buffer (RPMI 1640 + 1% FBS + 1% P.S). Subsequently, the Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line (Cat. GM-C30291) at a concentration of 1E5 cells/well was added, and the co-culture proceeded for an additional 7 hours. Firefly luciferase activity is then measured using the GMOne-Step Luciferase Reporter Gene Assay Kit (Cat. GM-040503). The results indicated a maximum blocking fold of approximately [172.2]. Data are shown by drug mass concentration.
Response to Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605). Serial dilutions of the Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605) (Cat. GM-59479AB) were incubated with 2E3 cells/well of the H_TNFSF15(TL1A) CHO-K1 Cell Line (Cat. GM-C19170) in a 96-well plate for 1 hour in assay buffer (RPMI 1640 + 1% FBS + 1% P.S). Subsequently, the Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line (Cat. GM-C30291) at a concentration of 1E5 cells/well was added, and the co-culture proceeded for an additional 6 hours. Firefly luciferase activity is then measured using the GMOne-Step Luciferase Reporter Gene Assay Kit (Cat. GM-040503). The results indicated a maximum blocking fold of approximately [26.0]. Data are shown by drug mass concentration.
Mouse_TNFRSF25(DR3) Reporter Jurkat Cell Line (Cat. GM-C30291) was determined by flow cytometry using Anti-TNFRSF25(DR3) hIgG1 Antibody(PTX-35) (Cat. GM-58913AB).
Specifications
Cat. NoGM-C30291
ProductMouse_TNFRSF25(DR3) Reporter Jurkat Cell Line
Product Format1 vial of frozen cells
Quantity5E6 Cells per vial,1 mL
Storage ConditionsLiquid nitrogen immediately upon receipt
Recovery MediumRPMI 1640+10% FBS+1% P.S
Growth mediumRPMI 1640+10% FBS+1% P.S+3.5 μg/mL Blasticidin+0.75 μg/mL Puromycin
NoteNone
Freezing Medium90% FBS+10% DMSO
Growth propertiesSuspension
Growth Conditions37°C, 5% CO₂
Safety considerationsBiosafety Level 2
Mycoplasma TestingThe cell line has been screened to confirm the absence of Mycoplasma species.
Materials
ReagentOrdering Information
BlasticidinGenomeditech/GM-040404
PuromycinGenomeditech/GM-040401
Pen/StrepThermo/15140-122
Fetal Bovine SerumCegrogen biotech/A0500-3010
RPMI 1640VivaCell/C3010-0500
Clear Flat-Bottom Immuno Nonsterile 96-Well PlatesThermo/442404
H_TNFSF15(TL1A) CHO-K1 Cell LineGenomeditech/GM-C19170
Anti-H_TNFSF15(TL1A) hIgG1 Antibody(PF-06480605)Genomeditech/GM-59479AB
Anti-TNFRSF25(DR3) hIgG1 Antibody(PTX-35)Genomeditech/GM-58913AB
GMOne-Step Luciferase Reporter Gene Assay KitGenomeditech/GM-040503
Cell Culture

Cell Recovery

Recovery Medium: RPMI 1640+10% FBS+1% P.S

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.

a)       Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 - 3 minutes).

b)       Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.

c)       Transfer the vial contents to a centrifuge tube containing 5.0 mL complete culture medium. And spin at approximately 176 x g for 5 minutes. Discard supernatant.

d)       Resuspend cell pellet with the recommended complete medium. And dispense the suspension into 1 - 2 T-25 culture flasks.

e)       Incubate the culture at 37°C in a suitable incubator. A 5% CO₂ in air atmosphere is recommended if using the medium described on this product sheet.

Cell Freezing

Freezing Medium: 90% FBS+10% DMSO

a)          Centrifuge at 176 x g for 3 minutes to collect cells.

b)         Resuspend the cells in pre-cooled freezing medium and adjust the cell density to 5E6 cells/mL.

c)          Aliquot 1 mL into each vial.

d)         Place the vial in a controlled-rate freezing container and store at -80°C for at least 1 day, then transfer to liquid nitrogen as soon as possible.

Cell passage

Growth medium: RPMI 1640+10% FBS+1% P.S+3.5 μg/mL Blasticidin+0.75 μg/mL Puromycin

Approximately 48-72 hours after the initial thawing, the cells can be passaged for the first time. After this initial passage, the culture medium can be adjusted to growth medium supplemented with antibiotics. If cells are not passaged within 48 hours, it is recommended to add some fresh recovery medium and place the flask horizontally.

a)          When the cell density reaches 1.5 - 2E6 cells/mL, subculture the cells. Do not allow the cell density to exceed 2E6 cells/mL.

b)         It is recommended to use T-25 flasks for subculturing.

c)          These cells are suspension cells, and it is recommended to use the "half-medium change" method to maintain optimal cell conditions during passaging.

d)         During passaging, you can directly add fresh growth medium to the culture flask, gently pipette to resuspend the cells, and then transfer the cell suspension to a new T-25 flask for continued culture.

Subcultivation Ratio: Maintain cultures at a cell concentraion between 3E5 and 1E6 viable cells/mL.

Medium Renewal: Every 2 to 3 days

Notes

a)          These cells are sensitive to density, so please ensure that the cell density is maintained within an appropriate range during culture and subculturing.

b)         During the first passage, pay attention to the nutrient supply; if not subculturing, make sure to add fresh recovery medium every other day as needed.

 

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