Andrographolide Inhibits Proliferation of Colon Cancer SW-480 Cells via Downregulating Notch Signaling Pathway
Background: Recently, Notch signaling pathway has gained attention as a potential therapeutic target for chemotherapeutic intervention. However, the efficacy of previously known inhibitors of Notch in colon cancer remains unclear. The purpose of this study was to determine the effect of andrographolide in aberrantly activated Notch signaling in SW-480 cells in vitro.
Methods: The potential cytostatic andrografolida on SW-480 cells was evaluated by 3- (4,5-dimethylthiazol-2-yl) -2, 5-diphenyl tetrazolium bromide (MTT) assay, assessment of morphology and colony formation test. FITC apoptosis activity was evaluated by Annexin V assay, 4 ‘, 6-diamidino-2-phenylindole (DAPI), Hoechst, Rhodamine 123 and Mito Tracker staining CMXRos. Scratching assay for assessment of potential migration. 7’-Dichlorodihydrofluorescein Diacetate (DCFH-DA) staining was used to evaluate Reactive Oxygen Species (ROS) generation. mRNA relative expression of Bax, Bcl-2, NOTCH 1 and jagged 1 was estimated by real-time quantitative reverse transcription PCR (qRT-PCR). cell cycle phase distribution was evaluated Annexin V-FITC / PI staining.
Results: MTT assay showed a dose and time dependent cytoxicity of andrographolide on SW-480 cells. It also inhibits migratory and colony forming potential of SW-480 cells. In addition, andrografolida also showed impaired mitochondrial membrane potential and induced apoptosis through nuclear condensation. Flow cytometric evaluation showed improved early and late andrographolide cells induced apoptosis and upregulation of proapoptotic (Bax and Bad) and downregulation of Bcl-2 antiapoptotic in SW-480 treated cells. Andrographolide plus intracellular ROS and induced G0 / G1 phase cell cycle arrest in SW480 colon cancer cells. Furthermore, andrografolida suppress Notch signaling by reducing the expression of NOTCH 1 and jagged one.
Conclusions: Our findings indicate that the andrographolide constraints SW-480 cell growth through inhibition of the Notch signaling pathway.
MicroRNA-133b exacerbate atherosclerosis by activating the Notch signaling pathway1
The purpose of this study was to determine the effect of miR-133b in atherosclerosis (AS). A rat model of AS (AS Group) was established, and serum levels of total cholesterol, triglycerides, high-density lipoprotein cholesterol and low-density lipoprotein (LDL) cholesterol were detected. The thoracic aorta tissue was subjected to hematoxylin and eosin staining for pathological examination. Mice were intravenously injected with microRNA (mir) -133b mimic (group miR-133b mimic + US) and miR-133b mimic negative control (miR-133b group NC + US).
Normal mice given the name of Sham group. reconstruction parameters of vascular, inflammatory factors Ratio Collagen / Vascular Area (CA / CVA) and serum of mice in each group were detected. MRNA expression was measured by quantitative reverse transcription-PCR and protein expression was determined by western blot analysis. An in vitro model of the US induced in vascular smooth muscle cells (VSMC) using oxidized (ox) -LDL. CCK-8 and wound healing tests used to detect cell proliferation and migration.
Compared with Sham group, the rats of the US group, AS + miR-133b mimic NC group and the group AS + miR-133b have a higher intima thickness (IT), tumor necrosis factor (TNF) -α and monocytes kemoatraktan protein (MCP) -1 level, as well as increased expression of Notch1 and Jagged1; and they have a lower medial thickness (MT), the ratio CA / CVA and Notch3 expression (all P <0.05).
) Hamster IgG, purified (Armenian, Isotype control) |
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| 20003-1AH | Alpha Diagnostics | 1 mg | EUR 263 |
 purified) G. Pig IgG Fc-Biotin conjugate (isotype control, non-immune) purified |
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| 20004-3-B | Alpha Diagnostics | 0.1 mg | EUR 225 |
) G. Pig IgG-Biotin conjugate (isotype control) |
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| 20004-B | Alpha Diagnostics | 100 ug | EUR 202 |
2 fragment, purified (isotype control)) Rat IgG F(ab')2 fragment, purified (isotype control) |
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| 20005-1-FAB2 | Alpha Diagnostics | 1 mg | EUR 202 |
-Biotin Conjuagte (isotype control), purified) Rat IgG (Fc)-Biotin Conjuagte (isotype control), purified |
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| 20005-1-FC-B | Alpha Diagnostics | 0.1 mg | EUR 225 |
) Rat IgG1-Biotin conjugate (isotype control) |
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| 20005-11-B | Alpha Diagnostics | 100 ug | EUR 202 |
) Rat IgG2a-APC conjugate (isotype control) |
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| 20005-12-APC | Alpha Diagnostics | 25 tests | EUR 202 |
) Rat IgG2a-Biotin conjugate (isotype control) |
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| 20005-12-B | Alpha Diagnostics | 100 ug | EUR 202 |
) Rat IgG2b-Biotin conjugate (isotype control) |
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| 20005-13-B | Alpha Diagnostics | 100 ug | EUR 202 |
) Rat IgG2c-Biotin conjugate (isotype control) |
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| 20005-14-B | Alpha Diagnostics | 100 ug | EUR 225 |
) Rat IgM-Biotin conjugate (isotype control) |
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| 20005-21-B | Alpha Diagnostics | 100 ug | EUR 225 |
) Rat IgG-Agarose conjugate(aff matrix) |
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| 20005-AS-1 | Alpha Diagnostics | 0.5 ml | EUR 164 |
 (Isotype control)) Rat IgG-Biotin conjugate (isotype control) (Isotype control) |
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| 20005-B | Alpha Diagnostics | 100 ug | EUR 164 |
, purified) Sheep IgG-Biotin Conjugate (isotype control, non-immune), purified |
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| 20006-1-B | Alpha Diagnostics | 0.5 mg | EUR 202 |
 control, purified) Sheep IgM-Biotin Conjugate (non-immune) control, purified |
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| 20006-2-B | Alpha Diagnostics | 0.1 mg | EUR 225 |
, purified) Sheep IgG Fc-Biotin Conjugate (isotype control, non-immune), purified |
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| 20006-4-B | Alpha Diagnostics | 0.5 mg | EUR 202 |
, purified) Human IgG-Biotin conjugate (isotype control, non-immune), purified |
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| 20007-1-B | Alpha Diagnostics | 0.5 mg | EUR 225 |
) Human IgG (>98%, non-immune, control, Liquid @ 10 mg/ml, azide free, bulk size) |
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| 20007-1-BL-1 | Alpha Diagnostics | 10 ml | Ask for price |
) Human IgG (>98%, non-immune, control, powder, azide free, bulk size) |
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| 20007-1-BP-1 | Alpha Diagnostics | 1 g | EUR 651 |
|
Human IgG (>98%, non-immune, control, powder, azide free, bulk size) |
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| 20007-1-BP-10 | Alpha Diagnostics | 10 g | EUR 4313 |
 Human IgG Fab fragment-Biotin Conjugate, purified |
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| 20007-1-FAB-B | Alpha Diagnostics | 0.1 mg | EUR 202 |
2 fragment, purified) Human IgG F(ab')2 fragment, purified |
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| 20007-1-FAB2 | Alpha Diagnostics | 1 mg | EUR 202 |
 Human IgG Fab2 fragment-Biotin Conjugate, purified |
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| 20007-1-FAB2-B | Alpha Diagnostics | 0.1 mg | EUR 202 |
, purified) Human IgG Fc-Biotin conjugate (isotype control, non-immune), purified |
|||
| 20007-1-Fc-B | Alpha Diagnostics | 0.1 mg | EUR 164 |
 fragment, purified (>95%, low endotoxin)) Human IgG (Fc) fragment, purified (>95%, low endotoxin) |
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| 20007-1-FC-LE | Alpha Diagnostics | 100 ul | EUR 225 |
 fragment-Biotin purified (>95%, low endotoxin)) Human IgG (Fc) fragment-Biotin purified (>95%, low endotoxin) |
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| 20007-1-FC-LE-B | Alpha Diagnostics | 25 ug | EUR 286 |
) Human IgG (serum origin, purified >97%, low endotoxin, azide free) |
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| 20007-1-LE-1 | Alpha Diagnostics | 1 mg | EUR 164 |
) Human IgG-Biotin (serum origin, purified >97%, low endotoxin, azide free) |
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| 20007-1-LE-BTN | Alpha Diagnostics | 100 ug | EUR 286 |
 TruStrip RDT Human IgG Rapid test cards, 10/pk |
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| 20007-1-RDT | Alpha Diagnostics | 1 Pk | EUR 171 |
 TruStrip RDT Human IgG Rapid test cards, 25/pk |
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| 20007-1-RDT-25 | Alpha Diagnostics | 1 Pk | EUR 293 |
) Human IgM, purified (native, isotype control, >96% pure, ELISA Grade) |
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| 20007-2 | Alpha Diagnostics | 1 mg | EUR 164 |
) Human IgM purified (isotype control, >96% pure, ELISA Grade) |
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| 20007-2-10 | Alpha Diagnostics | 10 mg | EUR 895 |
|
Human IgM purified (isotype control, >96% pure, ELISA Grade) |
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| 20007-2-5 | Alpha Diagnostics | 5 mg | EUR 529 |
) Human IgM (myeloma, >96%, non-immune, control; bulk size) |
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| 20007-2-B-100 | Alpha Diagnostics | 100 mg | Ask for price |
|
Human IgM (myeloma, >96%, non-immune, control; bulk size) |
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| 20007-2-B-1000 | Alpha Diagnostics | 1000 mg | Ask for price |
) Human IgM-Biotin conjugate (myeloma, isotype control) |
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| 20007-2-MB | Alpha Diagnostics | 100 ug | EUR 164 |
) Human IgM (Rheumatoid Factor plasma, >40,000 IU/ml, purifed for ELISA) |
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| 20007-2-RF | Alpha Diagnostics | 100 ul | EUR 469 |
, purified (isotype control, >95%, low endotoxin, azide free)) Human IgE (myeloma, kappa), purified (isotype control, >95%, low endotoxin, azide free) |
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| 20007-3-LE-50 | Alpha Diagnostics | 100 ug | EUR 408 |
, purified (isotype control, >95%, low endotoxin, azide free)) Human IgE-Biotin (myeloma, kappa), purified (isotype control, >95%, low endotoxin, azide free) |
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| 20007-3-LE-BTN | Alpha Diagnostics | 100 ul | EUR 408 |
-Biotin conjugate (isotype control)) Human IgM Fab mu fragment (myeloma)-Biotin conjugate (isotype control) |
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| 20007-4-B | Alpha Diagnostics | 100 ug | EUR 225 |
) Human IgG-Fc fragment (97%, low endotoxin, azide free) |
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| 20007-F-LE-100 | Alpha Diagnostics | 100 ug | EUR 164 |
) Human IgG-Fc fragment-Biotin (97%, low endotoxin, azide free) |
|||
| 20007-F-LE-BTN | Alpha Diagnostics | 100 ug | EUR 286 |
) Human IgG1-Biotin Conjugate (-ve control) |
|||
| 20007-G1-B | Alpha Diagnostics | 100 ug | EUR 164 |
 Human IgG1 Fc (103Cys/Ser, 99-330aa, His-tag, >95%, low endotoxin)) Recombinant (HEK) Human IgG1 Fc (103Cys/Ser, 99-330aa, His-tag, >95%, low endotoxin) |
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| 20007-G1-FC | Alpha Diagnostics | 50 ug | EUR 286 |
 Human IgG2-Biotin Conjugate |
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| 20007-G2-B | Alpha Diagnostics | 100 ug | EUR 164 |
 Human IgG2-Fc (257Ser/Ala, 99-327aa, His-tag, >95%, low endotoxin)) Recombinant (HEK) Human IgG2-Fc (257Ser/Ala, 99-327aa, His-tag, >95%, low endotoxin) |
|||
| 20007-G2-FC | Alpha Diagnostics | 50 ug | EUR 286 |
 Human IgG3-Biotin Conjugate |
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| 20007-G3-B | Alpha Diagnostics | 100 ug | EUR 164 |
 Human IgG3-Fc (99-377aa, His-tag, >95%, low endotoxin)) Recombinant (HEK) Human IgG3-Fc (99-377aa, His-tag, >95%, low endotoxin) |
|||
| 20007-G3-FC | Alpha Diagnostics | 50 ug | EUR 286 |
 Human IgG4-Biotin Conjugate |
|||
| 20007-G4-B | Alpha Diagnostics | 100 ug | EUR 164 |
 Human IgG4-Fc (108Ser/Pro, 99-327aa, His-tag, >95%, low endotoxin)) Recombinant (HEK) Human IgG4-Fc (108Ser/Pro, 99-327aa, His-tag, >95%, low endotoxin) |
|||
| 20007-G4-FC | Alpha Diagnostics | 50 ug | EUR 286 |
2 fragment, purified) Mouse IgG F(ab')2 fragment, purified |
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| 20008-1-Fab2 | Alpha Diagnostics | 1 mg | EUR 202 |
) Mouse IgG (>97%, ELISA/RIA/Diagnostic Grade; -ve mouse viruses & Protein A/G free) |
|||
| 20008-1-WA | Alpha Diagnostics | 1 mg | EUR 164 |
) Mouse IgG-Agarose conjugate (aff matrix) |
|||
| 20008-AS-1 | Alpha Diagnostics | 0.5 ml | EUR 164 |
) Mouse IgG-Biotin conjugate (isotype control) |
|||
| 20008-B | Alpha Diagnostics | 50 Tests | EUR 164 |
|
Mouse IgG-Biotin conjugate (isotype control) |
|||
| 20008-BT-1 | Alpha Diagnostics | 1 mg | EUR 164 |
) Mouse IgG Fab-Biotin conjugate (isotype control) |
|||
| 20008-Fab-B | Alpha Diagnostics | 100 ug | EUR 164 |
2-Biotin conjugate (isotype control)) Mouse IgG F(ab')2-Biotin conjugate (isotype control) |
|||
| 20008-Fab2-B | Alpha Diagnostics | 100 ug | EUR 164 |
2-FITC conjugate (isotype control)) Mouse IgG F(ab')2-FITC conjugate (isotype control) |
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| 20008-Fab2-F | Alpha Diagnostics | 100 ug | EUR 164 |
2-HRP conjugate (isotype control)) Mouse IgG F(ab')2-HRP conjugate (isotype control) |
|||
| 20008-Fab2-HP | Alpha Diagnostics | 100 ug | EUR 164 |
-Biotin conjugate (isotype control)) Mouse IgG F(c)-Biotin conjugate (isotype control) |
|||
| 20008-Fc-B | Alpha Diagnostics | 100 ug | EUR 164 |
 Rabbit IgG Fab-Biotin Conjugate, purified |
|||
| 20009-1-FAB-B | Alpha Diagnostics | 0.1 mg | EUR 202 |
2 fragment, purified) Rabbit IgG F(ab')2 fragment, purified |
|||
| 20009-1-FAB2 | Alpha Diagnostics | 1 mg | EUR 202 |
 Rabbit IgG (Fc) (101-323aa, 185Thr/Asn284Ser, ~33 kda, >95%, Low Endotoxin)) Recombinant (HEK) Rabbit IgG (Fc) (101-323aa, 185Thr/Asn284Ser, ~33 kda, >95%, Low Endotoxin) |
|||
| 20009-1-R-FC | Alpha Diagnostics | 50 ug | EUR 286 |
, purified) Rabbit IgM-Biotin Conjugate (isotype control, non-immune), purified |
|||
| 20009-2-1-B | Alpha Diagnostics | 0.1 mg | EUR 225 |
, purified) Rabbit IgA-Biotin Conjugate (isotype control, non-immune), purified |
|||
| 20009-3-1-B | Alpha Diagnostics | 0.1 mg | EUR 408 |
 Rabbit IgG-Agarose conjugate |
|||
| 20009-AS-1 | Alpha Diagnostics | 0.5 ml | EUR 164 |
-Biotin conjugate) Rabbit IgG (Whole, non-immune)-Biotin conjugate |
|||
| 20009-B | Alpha Diagnostics | 100 ug | EUR 164 |
) Rabbit IgG-biotinylated (isotype control) |
|||
| 20009-BT-1 | Alpha Diagnostics | 1 mg | EUR 286 |
2-biotin conjugate, isotype control) Rabbit IgG F(ab')2-biotin conjugate, isotype control |
|||
| 20009-FAb2-B | Alpha Diagnostics | 100 ug | EUR 164 |
2-FITC conjugate, isotype control) Rabbit IgG F(ab')2-FITC conjugate, isotype control |
|||
| 20009-FAb2-F | Alpha Diagnostics | 100 ug | EUR 164 |
2-HRP conjugate, isotype control) Rabbit IgG F(ab')2-HRP conjugate, isotype control |
|||
| 20009-FAb2-HP | Alpha Diagnostics | 100 ug | EUR 164 |
2-R-PE conjugate, isotype control) Rabbit IgG F(ab')2-R-PE conjugate, isotype control |
|||
| 20009-FAb2-PE | Alpha Diagnostics | 100 tests | EUR 225 |
-biotin conjugate, isotype control) Rabbit IgG F(c)-biotin conjugate, isotype control |
|||
| 20009-Fc-B | Alpha Diagnostics | 100 ug | EUR 164 |
, without azide, powder) Rabbit IgG, purified (serum non-immune, isotype control), without azide, powder |
|||
| 20009-WA-1 | Alpha Diagnostics | 1 mg | EUR 141 |
|
Rabbit IgG, purified (serum non-immune, isotype control), without azide, powder |
|||
| 20009-WA-5 | Alpha Diagnostics | 5 mg | EUR 286 |
 Human Insulin Antibody |
|||
| 20010-05011 | AssayPro | 150 ug | EUR 217 |
) Human Insulin Antibody (Biotin Conjugate) |
|||
| 20010-05021 | AssayPro | 150 ug | EUR 276 |
, purified) Chicken IgG-Biotin conjugate (isotype control, non-immune), purified |
|||
| 20010-1-B | Alpha Diagnostics | 0.5 mg | EUR 225 |
 purified) Chicken IgM-Biotin conjugate (isotype control, non-immune) purified |
|||
| 20010-2-1-b | Alpha Diagnostics | 0.1 mg | EUR 225 |
, purified) Chicken IgG-Fc-Biotin conjugate (isotype control, non-immune), purified |
|||
| 20010-3-B | Alpha Diagnostics | 0.1 mg | EUR 225 |
 Human Synaptobrevin-2 Antibody |
|||
| 20011-05011 | AssayPro | 150 ug | EUR 217 |
) Human Synaptobrevin-2 Antibody (Biotin Conjugate) |
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| 20011-05021 | AssayPro | 150 ug | EUR 276 |
) Human Synaptobrevin-2 AssayLite Antibody (FITC Conjugate) |
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| 20011-05041 | AssayPro | 150 ug | EUR 428 |
) Human Synaptobrevin-2 AssayLite Antibody (RPE Conjugate) |
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| 20011-05051 | AssayPro | 150 ug | EUR 428 |
) Human Synaptobrevin-2 AssayLite Antibody (APC Conjugate) |
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| 20011-05061 | AssayPro | 150 ug | EUR 428 |
) Human Synaptobrevin-2 AssayLite Antibody (PerCP Conjugate) |
|||
| 20011-05071 | AssayPro | 150 ug | EUR 471 |
 Goat IgG FAB fragment-Biotin conjugate |
|||
| 20011-1-FAB-B | Alpha Diagnostics | 100 tests | EUR 225 |
2 fragment, purified) Goat IgG F(ab')2 fragment, purified |
|||
| 20011-1-FAB2 | Alpha Diagnostics | 1 mg | EUR 164 |
) Goat IgG-Agarose conjugate(aff matrix) |
|||
| 20011-AS-1 | Alpha Diagnostics | 0.5 ml | EUR 164 |
-Biotin conjugate) Goat IgG (Whole, non-immune)-Biotin conjugate |
|||
| 20011-B | Alpha Diagnostics | 100 ug | EUR 164 |
2 fragment-Biotin conjugate) Goat IgG F(ab')2 fragment-Biotin conjugate |
|||
| 20011-FAB2-B | Alpha Diagnostics | 100 tests | EUR 202 |
2 fragment-Cy5 conjugate) Goat IgG F(ab')2 fragment-Cy5 conjugate |
|||
| 20011-FAB2-Cy5 | Alpha Diagnostics | 50 tests | EUR 202 |
2 fragment-FITC conjugate) Goat IgG F(ab')2 fragment-FITC conjugate |
|||
| 20011-FAB2-F | Alpha Diagnostics | 100 tests | EUR 202 |
2 fragment-HRP conjugate) Goat IgG FAB(ab')2 fragment-HRP conjugate |
|||
| 20011-FAB2-HP | Alpha Diagnostics | 100 tests | EUR 202 |
2 fragment-R-PE conjugate) Goat IgG F(ab')2 fragment-R-PE conjugate |
|||
| 20011-FAB2-PE | Alpha Diagnostics | 50 tests | EUR 202 |
-Biotin conjugate (isotype control)) Goat IgG (fc)-Biotin conjugate (isotype control) |
|||
| 20011-Fc-B | Alpha Diagnostics | 100 ug | EUR 164 |
In addition, miR-133b mimic promoted proliferation and migration, Notch1 and Jagged1 upregulated and downregulated Notch3 in VSMC ox-LDL-induced. Taken together, miR-133b worsen the US by activating Notch signaling pathway, which could serve as a potential target for the treatment of AS.
Anthony
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