CD253
Recombinant human CD253 used to assess chemoattractant properties of human trail on murine macrophages and smooth muscle cells in vitro
Image caption:
Chemoattractive Effects of TRAIL on Peritoneal Macrophages and Aortic SMCs
(a, c) Representative images of membranes stained with Diff-Quick solution. Peritoneal macrophages (arrows in panel a) and aortic SMCs (c) that have migrated to the membrane surface are seen in violet. Chambers were immersed in monocyte chemotactic protein-1 (MCP-1) (100 ng/ml), platelet-derived growth factor (PDGF) (0.5 μg/ml), or rh-TRAIL (1.0 μg/ml). The concentration of rh-OPG was 0.25–1.0 μg/ml. (b, d) Measurements of macrophages (b) or aortic SMCs (d) that have migrated per one mm2 of membrane area. Data are presented as mean ± SD (at least 3 independent experiments were performed); *p<0.05 or **p<0.01, as compared to controls or TRAIL alone.
From: Bumdelger B, Kokubo H, Kamata R, Fujii M, Yoshimura K, Aoki H, et al. (2016) Osteoprotegerin Prevents Development of Abdominal Aortic Aneurysms. PLoS ONE 11(1): e0147088.
Recombinant human CD253 used to assess induction of MMP-9 and Timp-1 by human trail on murine aortic smooth muscle cells in vitro
Image caption:
Induction of Mmp-9 and Timp-1 mRNA by Trail via the JNK or NF-κB Signaling Pathway. (a, b, d, e) Relative expression levels of Mmp-9 (a, b) and Timp-1 (d, e) mRNA in mouse aortic SMCs within a culture system induced by rh-TRAIL (10 ng/ml). SP-600125 (3–30 μM) (a) and AS-601245 (0.2–20 μM) (b) were used to inhibit the JNK signaling pathway. MG-132 (0.2–20 μM) (d) and BAY11-7082 (1–10 μM) (e) were used to inhibit the NF-κB pathway. Data are presented as mean ± SD; *p<0.05 or **p<0.01, as compared to controls or TRAIL alone. Samples were in triplicate for each group. (c, f) Representative double immunofluorescent staining images of abdominal aortas in wild-type and Opg-KO mice at six weeks, with pJNK (panels A-b, B-b, and C-b in c) or pNF-κB (panels D-b, E-b, and F-b in f) in green, and Mmp-9 (panels A-c, B-c, and C-c in c) or Timp-1 (panels D-c, E-c, and F-c in f) in red. pJNK signal merged with Mmp-9 (panels A-d, B-d, and C-d in c). pNF-κB signal merged with Timp-1 (panels A-d, B-d, and C-d in f). Rectangles in panels B and E show magnified areas in C and F, respectively. Nuclei are stained with DAPI in blue (m: medial layer; a: adventitia). Scale bars in panels A, B, D, and E represent 100 μm. Scale bars in panels C and F represent 5μμm.
From: Bumdelger B, Kokubo H, Kamata R, Fujii M, Yoshimura K, Aoki H, et al. (2016) Osteoprotegerin Prevents Development of Abdominal Aortic Aneurysms. PLoS ONE 11(1): e0147088.
Recombinant human CD253 (PHP125) used to stimulate mouse aortic smooth muscle cells in vitro
Image caption:
EPA prevents the up-regulation of Trail in Opg-KO mice via inhibition of JNK.
(A) Representative double immunofluorescent staining images for Trail (green) and Mmp-9 (red) are shown at six weeks after AAA induction in the aortas of wild-type and Opg-KO mice fed an EPA (+) diet. Rectangles with dotted lines in panels i-l and q-t are magnified in panels m-p and u-x, respectively. The border of the medial layer (m) with the intima or adventitia (a) is indicated by a white dotted line in panels m-p and u-x. Nuclei are stained with DAPI (blue). Scale bars represent 100 μm. (B-C) In the SMC culture system, Trail mRNA expression induced by rh-TRAIL (10 ng/mL) was inhibited by JNK inhibitors AS-601245 (0.2–20 μM) (B) and SP-600125 (3–30 μM) (C). Data are presented as mean ± SD; *p<0.05, as compared with controls or Trail alone. Experiments were independently repeated three times.
From: Kamata R, Bumdelger B, Kokubo H, Fujii M, Yoshimura K, Ishida T, et al. (2016)
EPA Prevents the Development of Abdominal Aortic Aneurysms through Gpr-120/Ffar-4.
PLoS ONE 11(10): e0165132.
Recombinant human CD253 (PHP125) used to stimulate mouse aortic smooth muscle cells in vitro
Image caption:
EPA reduces Trail-induced Mmp-9 expression via JNK.
(A) Western blotting analysis for pJNK/JNK from SMCs, which were cultured with rh-TRAIL (10 ng/mL) for 30 minutes with or without pre-incubation with EPA (0.1–10 μM). (B) Relative expression levels of Mmp-9 mRNA by qPCR from SMCs cultured with rh-TRAIL induction (10 ng/mL) for six hours with or without pre-incubation with EPA (0.1–10 μM). Data are presented as mean ± SD; *p<0.05, as compared with controls or Trail alone.
From: Kamata R, Bumdelger B, Kokubo H, Fujii M, Yoshimura K, Ishida T, et al. (2016)
EPA Prevents the Development of Abdominal Aortic Aneurysms through Gpr-120/Ffar-4.
PLoS ONE 11(10): e0165132.
Recombinant human CD253 (PHP125) used to stimulate mouse aortic smooth muscle cells in vitro
Image caption:
EPA reduces Mmp-9 expression by inactivating the Tak1-JNK pathway via activation of Gpr-120/Ffar-4.
(A) Representative double immunofluorescent staining image shows Gpr-120/Ffar-4 (green) and α-SMA (red) in the aortas of wild-type mice. Lower panels are magnified from upper panels. (B) Amplified Gpr-120/Ffar-4 mRNA samples derived from SMCs are shown. (C-F) Western blotting analysis for pTak-1/Tak-1 (C-E) and pJNK/JNK (F) from SMCs cultured for the indicated periods (C), 15 minutes (D and E), and 30 minutes (F) with rh-TRAIL (10 ng/mL). Phosphorylation of Tak-1 and JNK from SMCs, which were cultured with rh-TRAIL, was inhibited by EPA (D) and GW9508 (E and F). (G) Relative expression levels of Mmp-9 mRNA in SMCs cultured for six hours with rh-TRAIL (10 ng/mL). Expression of Mmp-9 in SMCs, induced by rh-TRAIL, were inhibited by GW9508. (H-J) Western blotting analysis for pTak-1/Tak-1 (H) and pJNK/JNK (I) from SMCs, which were treated with scrambled (NC) or Gpr-120/Ffar-4 siRNA (10 nM) and then cultured with rh-TRAIL (10 ng/mL) for 15 minutes (H) and 30 minutes (I). (J) Relative expression levels of Mmp-9 mRNA from SMCs treated with scrambled (NC) or Gpr-120/Ffar-4 siRNA (10 nM) and cultured for six hours with rh-TRAIL (10 ng/mL). Data are presented as mean ± SD; *p<0.05, as compared with controls or Trail alone.
From: Kamata R, Bumdelger B, Kokubo H, Fujii M, Yoshimura K, Ishida T, et al. (2016)
EPA Prevents the Development of Abdominal Aortic Aneurysms through Gpr-120/Ffar-4.
PLoS ONE 11(10): e0165132.
Recombinant human CD253 (PHP125) used to stimulate mouse aortic smooth muscle cells in vitro
Image caption:
siRNA-mediated knock-down of Gpr-120/Ffar-4 expression.
Relative expression levels of Gpr-120/Ffar-4 mRNA by qPCR from SMCs cultured with rh-TRAIL (10 mg/mL) for six hours with or without pre-incubation with EPA (10 μM). Gpr-120/Ffar-4 siRNA was applied 24 hours before pre-incubation with EPA. Data are presented as mean ± SD; *p<0.05, as compared with control.
From: Kamata R, Bumdelger B, Kokubo H, Fujii M, Yoshimura K, Ishida T, et al. (2016)
EPA Prevents the Development of Abdominal Aortic Aneurysms through Gpr-120/Ffar-4.
PLoS ONE 11(10): e0165132.
Recombinant human CD253 is a 19.6 kDa protein comprised of 168 amino acids which correspond to the TNF-homologous region of the extracellular domain of human TRAIL/ApoL2.
TNF-related apoptosis-inducing ligand (TRAIL) is expressed in many tissues including spleen, lung and prostate (Wiley et al, 1995) and is capable of inducing apoptosis in a tumour specific manner (Walczak et al, 1997). Apoptosis is initiated by the binding of the two death receptors DR4 and DR5 which initiate cell death signalling via the caspase cascade. There are however two decoy receptors DcR1 and DcR2 which competitively bind to CD253 but do not initiate apoptosis (Allen and El-Deiry, 2012).
A potential use for TRAIL as an anti-tumor therapeutic agent has been identified in numerous studies, demonstrating tumoricidal activity in approximately two thirds of tumor cell lines tested (Griffith et al, 1999).
Recombinant human CD253 is the immunogen used to produce Rabbit anti human CD253 (AHP1215).
TNF-related apoptosis-inducing ligand (TRAIL) is expressed in many tissues including spleen, lung and prostate (Wiley et al, 1995) and is capable of inducing apoptosis in a tumour specific manner (Walczak et al, 1997). Apoptosis is initiated by the binding of the two death receptors DR4 and DR5 which initiate cell death signalling via the caspase cascade. There are however two decoy receptors DcR1 and DcR2 which competitively bind to CD253 but do not initiate apoptosis (Allen and El-Deiry, 2012).
A potential use for TRAIL as an anti-tumor therapeutic agent has been identified in numerous studies, demonstrating tumoricidal activity in approximately two thirds of tumor cell lines tested (Griffith et al, 1999).
Recombinant human CD253 is the immunogen used to produce Rabbit anti human CD253 (AHP1215).
Product Details
- Target Species
- Human
- Species Cross-Reactivity
-
Target Species Cross Reactivity Mouse - N.B. Antibody reactivity and working conditions may vary between species.
- Product Form
- Purified recombinant protein - lyophilised
- Reconstitution
- Reconstitute with 0.5ml distilled water. For extended storage, the addition of 0.1% BSA is recommended.
Care should be taken during reconstitution as the protein may appear as a film at the bottom of the vial. Bio-Rad recommend that the vial is gently mixed after reconstitution. - Preparation
- Recombinant protein expressed in E.coli
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- None present
- Purity
- >98% by SDS PAGE and HPLC analysis
- Approx. Protein Concentrations
- 0.1mg/ml after reconstitution.
- Protein Molecular Weight
- 19.6kD (169 amino acid sequence)
- Endotoxin Level
- <0.1ng/ug
Storage Information
- Storage
- Prior to reconstitution store at +4oC. Following reconstitution store at -20oC.
This product should be stored undiluted.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the protein. Should this product contain a precipitate we recommend microcentrifugation before use. - Shelf Life
- 3 months from date of reconstitution.
More Information
- UniProt
- P50591 Related reagents
- Entrez Gene
- TNFSF10 Related reagents
- GO Terms
- GO:0007165 signal transduction
- GO:0005887 integral to plasma membrane
- GO:0005125 cytokine activity
- GO:0005164 tumor necrosis factor receptor binding
- GO:0046872 metal ion binding
- GO:0005615 extracellular space
- GO:0005625 soluble fraction
- GO:0006919 activation of caspase activity
- GO:0006955 immune response
- GO:0007267 cell-cell signaling
- GO:0008624 induction of apoptosis by extracellular signals
- GO:0008633 activation of pro-apoptotic gene products
- GO:0043123 positive regulation of I-kappaB kinase/NF-kappaB cascade
- Regulatory
- For research purposes only
Applications of CD253
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA | 0.2 | 0.4ng/well | |
| Functional Assays | 0.2 | 20ng/ml | |
| Western Blotting | 1.5 | 3.0ng/lane |
Where this protein has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the protein for use in their own system using appropriate positive/negative controls.
- ELISA
- Recombinant human CD253 may be used as a standard for ELISA applications with anti human CD253 (purified)
- Western Blotting
- Recombinant human CD253 may be used as the positive control for western blotting applications with anti human CD253 (purified)
Copyright © 2018 Bio-Rad Antibodies (formerly AbD Serotec)
Product Specific References
References for CD253
-
Bumdelger, B. et al. (2016) Osteoprotegerin Prevents Development of Abdominal Aortic Aneurysms.
PLoS One. 11 (1): e0147088. -
Kamata, R. et al. (2016) EPA Prevents the Development of Abdominal Aortic Aneurysms through Gpr-120/Ffar-4.
PLoS One. 11 (10): e0165132.
