mouse anti human cd40 monoclonal antibody lob7 6 (Bio-Rad)

Bio-Rad is a verified supplier

Bio-Rad manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Bio-Rad mouse anti human cd40 monoclonal antibody lob7 6

FIG. 1. Induction of cytokines in FSDCs and BMDCs during culture with <t>anti-CD40</t> mAb 3/23. mRNA was isolated from FSDCs or BMDCs which were either incubated with 30 g/ml anti-CD40 mAb 3/23 or left untreated for 24 h. The mRNA was used to obtain first strand cDNA, which was used as a template in RT-PCR reactions using protocols described under “Materials and Methods.” Utilizing this method, cDNA species encoding murine IL-6, IL-12 (p35 and p40), IFN, IL-4, and -actin were amplified. For IFN and IL-4, positive control RT-PCR reactions are shown on the far left-hand side and used mRNA extracted from total blood lymphocytes (TBL) taken from a Balb/c mouse. The gels shown are representative of at least two inde- pendent experiments. A 1-kilobase DNA ladder was run alongside the PCR products to confirm correct sizes of the amplified cDNA fragments.

Mouse Anti Human Cd40 Monoclonal Antibody Lob7 6, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+human+cd40+monoclonal+antibody+lob7+6/Mouse+anti+Human+CD40/10__1074_slash_jbc__m109250200-68-0-9
Average 93 stars, based on 70 article reviews

mouse anti human cd40 monoclonal antibody lob7 6 - by Bioz Stars, 2026-09

93/100 stars

Images

1) Product Images from "CD40 Induces Interleukin-6 Gene Transcription in Dendritic Cells"

Article Title: CD40 Induces Interleukin-6 Gene Transcription in Dendritic Cells

Journal: Journal of Biological Chemistry

doi: 10.1074/jbc.m109250200

FIG. 1. Induction of cytokines in FSDCs and BMDCs during culture with anti-CD40 mAb 3/23. mRNA was isolated from FSDCs or BMDCs which were either incubated with 30 g/ml anti-CD40 mAb 3/23 or left untreated for 24 h. The mRNA was used to obtain first strand cDNA, which was used as a template in RT-PCR reactions using protocols described under “Materials and Methods.” Utilizing this method, cDNA species encoding murine IL-6, IL-12 (p35 and p40), IFN, IL-4, and -actin were amplified. For IFN and IL-4, positive control RT-PCR reactions are shown on the far left-hand side and used mRNA extracted from total blood lymphocytes (TBL) taken from a Balb/c mouse. The gels shown are representative of at least two inde- pendent experiments. A 1-kilobase DNA ladder was run alongside the PCR products to confirm correct sizes of the amplified cDNA fragments.

Figure Legend Snippet: FIG. 1. Induction of cytokines in FSDCs and BMDCs during culture with anti-CD40 mAb 3/23. mRNA was isolated from FSDCs or BMDCs which were either incubated with 30 g/ml anti-CD40 mAb 3/23 or left untreated for 24 h. The mRNA was used to obtain first strand cDNA, which was used as a template in RT-PCR reactions using protocols described under “Materials and Methods.” Utilizing this method, cDNA species encoding murine IL-6, IL-12 (p35 and p40), IFN, IL-4, and -actin were amplified. For IFN and IL-4, positive control RT-PCR reactions are shown on the far left-hand side and used mRNA extracted from total blood lymphocytes (TBL) taken from a Balb/c mouse. The gels shown are representative of at least two inde- pendent experiments. A 1-kilobase DNA ladder was run alongside the PCR products to confirm correct sizes of the amplified cDNA fragments.

Techniques Used: Isolation, Incubation, Reverse Transcription Polymerase Chain Reaction, Amplification, Positive Control

FIG. 2. Induction of IL-6 promoter activity by anti-CD40 and requirement for specific regulatory DNA motifs. A, FSDCs were transfected with 100 ng of pRLTK and 1 g of either wild type pIL6- Luc651 or mutated pIL6-Luc651 constructs carrying mutation in the AP1, NF-B, and NF-IL6 sites. The transfected cultures were split into two flasks. 24 h later, one culture flask was incubated with 30 g/ml anti-CD40 mAb 3/23 for a further 24 h, while the remaining flask was left untreated. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated samples are in white. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Statistical analysis was performed by Student’s t test. *, **, and *** denote p 0.05, 0.01, and 0.005, respectively. B, fold induction of the sample incubated with anti-CD40 mAb 3/23 versus the non-treated sample for each of the pIL6-Luc651 constructs.

Figure Legend Snippet: FIG. 2. Induction of IL-6 promoter activity by anti-CD40 and requirement for specific regulatory DNA motifs. A, FSDCs were transfected with 100 ng of pRLTK and 1 g of either wild type pIL6- Luc651 or mutated pIL6-Luc651 constructs carrying mutation in the AP1, NF-B, and NF-IL6 sites. The transfected cultures were split into two flasks. 24 h later, one culture flask was incubated with 30 g/ml anti-CD40 mAb 3/23 for a further 24 h, while the remaining flask was left untreated. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated samples are in white. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Statistical analysis was performed by Student’s t test. *, **, and *** denote p 0.05, 0.01, and 0.005, respectively. B, fold induction of the sample incubated with anti-CD40 mAb 3/23 versus the non-treated sample for each of the pIL6-Luc651 constructs.

Techniques Used: Activity Assay, Transfection, Construct, Mutagenesis, Incubation, Luciferase

FIG. 3. Induction of the IL-6 pro- moter requires activation of NF-B. A, FSDCs were transfected with 100 ng of pRLTK, 1 g of wild type pIL6-Luc651, and 2 g of IB trans-dominant negative (IBD) expression vector or pcDNA3 as control. The transfected cultures were split into two flasks. 24 h after transfec- tion, one flask was incubated with 30 g/ml anti-CD40 mAb 3/23 for 24 h while the remaining flask was left untreated. Luciferase activity was determined 48 h after transfection. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated sam- ples are in white. Statistical analysis was performed by Student’s t test. *** denotes p 0.005. B, fold induction of the samples incubated with anti-CD40 mAb 3/23 ver- sus the non-treated samples as shown in Fig. 3A. C, mRNA was isolated from BM- DCs which were either untreated () or incubated for 24 h with 30 g/ml anti- CD40 mAb 3/23 together with either 5 M MG132 dissolved in Me2SO or Me2SO alone. The mRNA was used to obtain first strand cDNA, which was used as a tem- plate in RT-PCR. cDNA species encoding murine IL-6 and -actin were amplified over 32 and 27 cycles in RT-PCR reactions using protocols described under “Materi- als and Methods.” The gels shown are rep- resentative of at least two independent experiments. A 1-kilobase DNA ladder was run alongside the PCR products to confirm correct sizes of the amplified cDNA fragments.

Figure Legend Snippet: FIG. 3. Induction of the IL-6 pro- moter requires activation of NF-B. A, FSDCs were transfected with 100 ng of pRLTK, 1 g of wild type pIL6-Luc651, and 2 g of IB trans-dominant negative (IBD) expression vector or pcDNA3 as control. The transfected cultures were split into two flasks. 24 h after transfec- tion, one flask was incubated with 30 g/ml anti-CD40 mAb 3/23 for 24 h while the remaining flask was left untreated. Luciferase activity was determined 48 h after transfection. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated sam- ples are in white. Statistical analysis was performed by Student’s t test. *** denotes p 0.005. B, fold induction of the samples incubated with anti-CD40 mAb 3/23 ver- sus the non-treated samples as shown in Fig. 3A. C, mRNA was isolated from BM- DCs which were either untreated () or incubated for 24 h with 30 g/ml anti- CD40 mAb 3/23 together with either 5 M MG132 dissolved in Me2SO or Me2SO alone. The mRNA was used to obtain first strand cDNA, which was used as a tem- plate in RT-PCR. cDNA species encoding murine IL-6 and -actin were amplified over 32 and 27 cycles in RT-PCR reactions using protocols described under “Materi- als and Methods.” The gels shown are rep- resentative of at least two independent experiments. A 1-kilobase DNA ladder was run alongside the PCR products to confirm correct sizes of the amplified cDNA fragments.

Techniques Used: Activation Assay, Transfection, Dominant Negative Mutation, Expressing, Plasmid Preparation, Control, Incubation, Luciferase, Activity Assay, Isolation, Reverse Transcription Polymerase Chain Reaction, Amplification

FIG. 4. EMSA analysis of NF-B and CBF1 in anti-CD40 stimulated FSDC. A, nuclear extracts from control or FSDC cells treated with anti-mCD40 mAb 3/23 for 24 h were isolated and 2 g used in EMSA with NF-B double stranded oligonucleotide probe. Two (1 and 2) specific DNA-protein complexes were assembled and are denoted by arrows. Supershift analysis was performed on control and treated samples using antisera recognizing p50 and p65 or JunB as a control. B, 2 g of nuclear extracts from FSDC cells treated with anti-mCD40 mAb 3/23 for 24 h (as obtained in A) were incubated with double stranded NF-B oligonucleotide probe, or NF-B probe lacking p50/p65 or CBF1-binding sites. The nucleotide substitutions introduced into the wild type NF-B oligonucleotide to generate mutant oligonucleotides lacking p50/p65 or CBF1-binding sites are shown below the EMSA gel. C, 2 g of nuclear extracts from FSDC cells treated with anti-mCD40 mAb 3/23 for 24 h (as obtained in A) were incubated with double stranded NF-B oligonucleotide probe and supershift analysis performed using antisera recognizing p50, p65, CBF1, or JunB as a control.

Figure Legend Snippet: FIG. 4. EMSA analysis of NF-B and CBF1 in anti-CD40 stimulated FSDC. A, nuclear extracts from control or FSDC cells treated with anti-mCD40 mAb 3/23 for 24 h were isolated and 2 g used in EMSA with NF-B double stranded oligonucleotide probe. Two (1 and 2) specific DNA-protein complexes were assembled and are denoted by arrows. Supershift analysis was performed on control and treated samples using antisera recognizing p50 and p65 or JunB as a control. B, 2 g of nuclear extracts from FSDC cells treated with anti-mCD40 mAb 3/23 for 24 h (as obtained in A) were incubated with double stranded NF-B oligonucleotide probe, or NF-B probe lacking p50/p65 or CBF1-binding sites. The nucleotide substitutions introduced into the wild type NF-B oligonucleotide to generate mutant oligonucleotides lacking p50/p65 or CBF1-binding sites are shown below the EMSA gel. C, 2 g of nuclear extracts from FSDC cells treated with anti-mCD40 mAb 3/23 for 24 h (as obtained in A) were incubated with double stranded NF-B oligonucleotide probe and supershift analysis performed using antisera recognizing p50, p65, CBF1, or JunB as a control.

Techniques Used: Control, Isolation, Incubation, Binding Assay, Mutagenesis

FIG. 5. Induction of AP-1 activity by CD40 ligation. A, nuclear extracts from control or FSDC cells treated with anti-mCD40 mAb 3/23 for 16 h were isolated and 10 g used in EMSA with AP1 double stranded oligonucleotide probe in presence of 100 M excess of unlabeled nonspecific oligonucleotide (Sp1) or unlabeled specific AP1 oligonucleotide. B, supershift analysis was performed on 3/23 mAb treated FSDC samples using antisera recognizing c-Jun, JunD, JunB, c-Fos, or Sp1 as a control. Supershift complexes are shown for extracts incubated with JunB and JunD antisera. Asterisks placed to the left of the supershift complexes are included to aid identification of these species. C, immunoblot analysis of JunD, JunB, and c-Jun protein expression was performed on crude cytoplasmic and nuclear extracts from control and FSDCs treated with 3/23 mAb for 16 h. All gels are representative of three independent experiments. D, FSDCs were transfected with 100 ng of pRLTK, 1 g of wild type pIL6-Luc651, and 2 g of JunD dominant negative expression vector pRSV-JunD or pRSV as control. The transfected cultures were split into two flasks and after 24 h one flask was incubated with 30 g/ml anti-CD40 mAb 3/23 while the remaining flask was left untreated. Luciferase activity was determined 48 h after transfection. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated samples are in white. Statistical analysis was performed by Student’s t test. ** denotes p 0.01.

Figure Legend Snippet: FIG. 5. Induction of AP-1 activity by CD40 ligation. A, nuclear extracts from control or FSDC cells treated with anti-mCD40 mAb 3/23 for 16 h were isolated and 10 g used in EMSA with AP1 double stranded oligonucleotide probe in presence of 100 M excess of unlabeled nonspecific oligonucleotide (Sp1) or unlabeled specific AP1 oligonucleotide. B, supershift analysis was performed on 3/23 mAb treated FSDC samples using antisera recognizing c-Jun, JunD, JunB, c-Fos, or Sp1 as a control. Supershift complexes are shown for extracts incubated with JunB and JunD antisera. Asterisks placed to the left of the supershift complexes are included to aid identification of these species. C, immunoblot analysis of JunD, JunB, and c-Jun protein expression was performed on crude cytoplasmic and nuclear extracts from control and FSDCs treated with 3/23 mAb for 16 h. All gels are representative of three independent experiments. D, FSDCs were transfected with 100 ng of pRLTK, 1 g of wild type pIL6-Luc651, and 2 g of JunD dominant negative expression vector pRSV-JunD or pRSV as control. The transfected cultures were split into two flasks and after 24 h one flask was incubated with 30 g/ml anti-CD40 mAb 3/23 while the remaining flask was left untreated. Luciferase activity was determined 48 h after transfection. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated samples are in white. Statistical analysis was performed by Student’s t test. ** denotes p 0.01.

Techniques Used: Activity Assay, Ligation, Control, Isolation, Incubation, Western Blot, Expressing, Transfection, Dominant Negative Mutation, Plasmid Preparation, Luciferase

FIG. 6. Requirement for specific amino acid sequences in the cytoplasmic domain of CD40. A, 1 g of wild type pIL6–651Luc was transfected into FSDC cells along with 100 ng of pRLTK and 2 g of empty vector pcDNA3 or pcDNA3-derived expression vectors carrying cDNA cassettes for wild type hCD40, truncated hCD40 containing three remaining intracellular residues (hCD40KKV), hCD40 mutants carrying point mutations (hCD40T254A, hCD40T254E, and hCD40T254S), hCD40 with a point mutation and a deletion of carboxyl-terminal 15 amino acids (hCD40T254A262), hCD40 with 15 carboxyl-terminal amino acids deleted (hCD40262), or a fusion protein made up of hCD450 extracellular and transmembrane domains and hCD22 intracellular domain. The transfected cultures were split into two flasks, one flask was treated with 300 g/ml anti-human CD40 mAb LOB7.6 for 24 h and the other was left untreated. 24 h later, cells were harvested and luciferase assays performed. Samples treated with LOB7.6 are depicted in black solid boxes, whereas untreated samples are in white. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of six independent transfection experiments. B, FSDC cells were transfected with 1 g of IB-Luc, 100 ng of pRLTK and hCD40 expression vectors and treated as already described in Fig. 5A. Luciferase activities were normalized to pRLTK activity and expressed as the means S.E. of five independent transfection experiments.

Figure Legend Snippet: FIG. 6. Requirement for specific amino acid sequences in the cytoplasmic domain of CD40. A, 1 g of wild type pIL6–651Luc was transfected into FSDC cells along with 100 ng of pRLTK and 2 g of empty vector pcDNA3 or pcDNA3-derived expression vectors carrying cDNA cassettes for wild type hCD40, truncated hCD40 containing three remaining intracellular residues (hCD40KKV), hCD40 mutants carrying point mutations (hCD40T254A, hCD40T254E, and hCD40T254S), hCD40 with a point mutation and a deletion of carboxyl-terminal 15 amino acids (hCD40T254A262), hCD40 with 15 carboxyl-terminal amino acids deleted (hCD40262), or a fusion protein made up of hCD450 extracellular and transmembrane domains and hCD22 intracellular domain. The transfected cultures were split into two flasks, one flask was treated with 300 g/ml anti-human CD40 mAb LOB7.6 for 24 h and the other was left untreated. 24 h later, cells were harvested and luciferase assays performed. Samples treated with LOB7.6 are depicted in black solid boxes, whereas untreated samples are in white. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of six independent transfection experiments. B, FSDC cells were transfected with 1 g of IB-Luc, 100 ng of pRLTK and hCD40 expression vectors and treated as already described in Fig. 5A. Luciferase activities were normalized to pRLTK activity and expressed as the means S.E. of five independent transfection experiments.

Techniques Used: Transfection, Plasmid Preparation, Derivative Assay, Expressing, Mutagenesis, Luciferase, Activity Assay

FIG. 7. Inhibition of anti-CD40 induced IL-6 promoter activity by dominant negative TRAF2. A, mRNA was isolated from FSDCs which were either incubated with 30 g/ml anti-CD40 mAb 3/23, or left untreated for 24 h. The mRNA was used to obtain first strand cDNA, which was used as a template in RT-PCR reactions using protocols described under “Materials and Methods.” Utilizing this method, cDNA species encoding murine TRAF2, TRAF6, and -actin were amplified over 35, 35, and 28 PCR cycles, respectively. The gels shown are representative of at least two independent experiments. A 1-kilobase DNA ladder was run alongside the PCR products to confirm correct sizes of the amplified cDNA fragments. B, FSDCs were transfected with 100 ng of pRLTK, 1 g of IB-Luc, and 2 g of TRAF2 dominant negative (TRAF2 DN) expression vector or empty control vector. The cultures were split into 2 and 24 h after the transfection one-half was incubated with 30 g/ml anti-CD40 mAb 3/23 for 24 h. Luciferase activity was determined 48 h after transfection. C, FSDCs were transfected with 100 ng of pRLTK, 1 g of IB-Luc, and 2 g of TRAF6 dominant negative (TRAF6 DN) and processed as described for B. For both B and C, luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated samples are in white. Statistical analysis was performed by Student’s t test. *** denotes p 0.005.

Figure Legend Snippet: FIG. 7. Inhibition of anti-CD40 induced IL-6 promoter activity by dominant negative TRAF2. A, mRNA was isolated from FSDCs which were either incubated with 30 g/ml anti-CD40 mAb 3/23, or left untreated for 24 h. The mRNA was used to obtain first strand cDNA, which was used as a template in RT-PCR reactions using protocols described under “Materials and Methods.” Utilizing this method, cDNA species encoding murine TRAF2, TRAF6, and -actin were amplified over 35, 35, and 28 PCR cycles, respectively. The gels shown are representative of at least two independent experiments. A 1-kilobase DNA ladder was run alongside the PCR products to confirm correct sizes of the amplified cDNA fragments. B, FSDCs were transfected with 100 ng of pRLTK, 1 g of IB-Luc, and 2 g of TRAF2 dominant negative (TRAF2 DN) expression vector or empty control vector. The cultures were split into 2 and 24 h after the transfection one-half was incubated with 30 g/ml anti-CD40 mAb 3/23 for 24 h. Luciferase activity was determined 48 h after transfection. C, FSDCs were transfected with 100 ng of pRLTK, 1 g of IB-Luc, and 2 g of TRAF6 dominant negative (TRAF6 DN) and processed as described for B. For both B and C, luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated samples are in white. Statistical analysis was performed by Student’s t test. *** denotes p 0.005.

Techniques Used: Inhibition, Activity Assay, Dominant Negative Mutation, Isolation, Incubation, Reverse Transcription Polymerase Chain Reaction, Amplification, Transfection, Expressing, Plasmid Preparation, Control, Luciferase

FIG. 8. Induction of IL-6 gene transcription by anti-CD40 is a transient response. mRNA was isolated from control or FSDCs treated with anti-mCD40 mAb 3/23 for 24, 48, or 72 h. mRNA was used to obtain first strand cDNA which was used as a template in RT-PCR reactions using protocols described under “Materials and Methods.” Utilizing this method, cDNA species encoding murine IL-6 and -actin were amplified. The gels shown are representative of at least two independent experiments. A 1-kilobase DNA ladder was run alongside the PCR products to confirm correct sizes of the amplified cDNA fragments.

Figure Legend Snippet: FIG. 8. Induction of IL-6 gene transcription by anti-CD40 is a transient response. mRNA was isolated from control or FSDCs treated with anti-mCD40 mAb 3/23 for 24, 48, or 72 h. mRNA was used to obtain first strand cDNA which was used as a template in RT-PCR reactions using protocols described under “Materials and Methods.” Utilizing this method, cDNA species encoding murine IL-6 and -actin were amplified. The gels shown are representative of at least two independent experiments. A 1-kilobase DNA ladder was run alongside the PCR products to confirm correct sizes of the amplified cDNA fragments.

Techniques Used: Isolation, Control, Reverse Transcription Polymerase Chain Reaction, Amplification

FIG. 9. Prolonged induction of CBF1 DNA binding activity and repression of IL-6 promoter function. A, 2 g of nuclear extract isolated from FSDC cells treated with anti-mCD40 mAb 3/23 for 0, 2, 4, 6, 16, 24, and 48 h was used in EMSA with NF-B double stranded oligonucleotide probe. B, FSDCs were transfected with 100 ng of pRLTK, 1 g of wild type pIL6-Luc651, and 2 g of CBF1 expression vector pJH282 or empty vector pSG5. The cultures were split into two flasks and 24 h after transfection one flask was incubated with 30 g/ml anti-CD40 mAb 3/23 for 24 h, while the remaining flask was left untreated. The cells were then harvested and luciferase assay performed. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated samples are in white. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Statistical analysis was performed by Student’s t test. *** denotes p 0.005.

Figure Legend Snippet: FIG. 9. Prolonged induction of CBF1 DNA binding activity and repression of IL-6 promoter function. A, 2 g of nuclear extract isolated from FSDC cells treated with anti-mCD40 mAb 3/23 for 0, 2, 4, 6, 16, 24, and 48 h was used in EMSA with NF-B double stranded oligonucleotide probe. B, FSDCs were transfected with 100 ng of pRLTK, 1 g of wild type pIL6-Luc651, and 2 g of CBF1 expression vector pJH282 or empty vector pSG5. The cultures were split into two flasks and 24 h after transfection one flask was incubated with 30 g/ml anti-CD40 mAb 3/23 for 24 h, while the remaining flask was left untreated. The cells were then harvested and luciferase assay performed. Samples treated with 3/23 are depicted in black solid boxes, whereas untreated samples are in white. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Statistical analysis was performed by Student’s t test. *** denotes p 0.005.

Techniques Used: Binding Assay, Activity Assay, Isolation, Transfection, Expressing, Plasmid Preparation, Incubation, Luciferase

FIG. 10. Conservation of CD40 signaling events between two distinct mouse DC lines (FSDC and DC2.4). To confirm the generality of findings with FSDC a series of key experiments were repeated using the mouse DC line DC2.4. DC2.4s were transfected with 100 ng of pRLTK, 1 g of wild type pIL6-Luc651wt; A, 2 g of IB dominant negative (IB DN) expression vector or pcDNA3 empty vector as a control; B, 2 g of JunD dominant negative (pRSV JunD) expression vector or pRSV empty vector as a control; C, 2 g of TRAF2 dominant negative (TRAF2 DN) expression vector or pRK5 empty vector as a control; D, 2 g of TRAF6 dominant negative (TRAF6 DN) expression vector or pcDNA3 empty vector as a control; and E, 2 g of CBF1 dominant negative (pJH282) expression vector or pSG5 empty vector as a control. The transfected cultures were split into two flasks. 24 h after transfection, one flask was incubated with 30 g/ml anti-CD40 mAb 3/23 for 24 h while the remaining flask was left untreated. Luciferase activity was determined 48 h after transfection. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Samples treated with 3/23 are depicted in solid black boxes, whereas untreated samples are in white. Statistical analysis was performed by Student’s t test. * and *** denote p 0.05 and p 0.005, respectively.

Figure Legend Snippet: FIG. 10. Conservation of CD40 signaling events between two distinct mouse DC lines (FSDC and DC2.4). To confirm the generality of findings with FSDC a series of key experiments were repeated using the mouse DC line DC2.4. DC2.4s were transfected with 100 ng of pRLTK, 1 g of wild type pIL6-Luc651wt; A, 2 g of IB dominant negative (IB DN) expression vector or pcDNA3 empty vector as a control; B, 2 g of JunD dominant negative (pRSV JunD) expression vector or pRSV empty vector as a control; C, 2 g of TRAF2 dominant negative (TRAF2 DN) expression vector or pRK5 empty vector as a control; D, 2 g of TRAF6 dominant negative (TRAF6 DN) expression vector or pcDNA3 empty vector as a control; and E, 2 g of CBF1 dominant negative (pJH282) expression vector or pSG5 empty vector as a control. The transfected cultures were split into two flasks. 24 h after transfection, one flask was incubated with 30 g/ml anti-CD40 mAb 3/23 for 24 h while the remaining flask was left untreated. Luciferase activity was determined 48 h after transfection. Luciferase activities were normalized to pRLTK activity and expressed as the mean S.E. of three independent transfection experiments. Samples treated with 3/23 are depicted in solid black boxes, whereas untreated samples are in white. Statistical analysis was performed by Student’s t test. * and *** denote p 0.05 and p 0.005, respectively.

Techniques Used: Transfection, Dominant Negative Mutation, Expressing, Plasmid Preparation, Control, Incubation, Luciferase, Activity Assay

Isolation:

Article Title: CD40 Induces Interleukin-6 Gene Transcription in Dendritic Cells
Article Snippet: CD40 Antibodies—Rat monoclonal antibody 3/23 was raised against mouse CD40 (31).CD40 Antibodies—Rat monoclonal antibody 3/23 was raised against mouse CD40 (31).. Mouse anti-human CD40 monoclonal antibody LOB7.6 was obtained from Serotec, UK.. Plasmid DNA—All plasmid DNA was prepared using a commercial DNA extraction and isolation kit (Maxiprep, Qiagen).Plasmid DNA—All plasmid DNA was prepared using a commercial DNA extraction and isolation kit (Maxiprep, Qiagen).

Review

mouse anti human cd40 monoclonal antibody lob7 6 (Bio-Rad)

Structured Review

Images

1) Product Images from "CD40 Induces Interleukin-6 Gene Transcription in Dendritic Cells"

Related Articles

Isolation:

Incubation:

Reverse Transcription Polymerase Chain Reaction:

Amplification:

Positive Control:

Activity Assay:

Transfection:

Construct:

Mutagenesis:

Luciferase:

Activation Assay:

Dominant Negative Mutation:

Expressing:

Plasmid Preparation:

Control:

Binding Assay:

Ligation:

Western Blot:

Derivative Assay:

Inhibition:

Similar Products

Image Search Results