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Santa Cruz Biotechnology specic primary antibodies against timp3

Fig. 3 <t>TIMP3</t> was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 30-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. **P < 0.01 or ***P < 0.001 vs. respective control.

Specic Primary Antibodies Against Timp3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3."

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3.

Journal: RSC advances

doi: 10.1039/c8ra10552a

Fig. 3 TIMP3 was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 30-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. **P < 0.01 or ***P < 0.001 vs. respective control.

Figure Legend Snippet: Fig. 3 TIMP3 was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 30-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. **P < 0.01 or ***P < 0.001 vs. respective control.

Techniques Used: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Construct, Transfection, Quantitative RT-PCR, Western Blot, Control

Fig. 4 TGF-b1 repressed the expression of TIMP3. HaCaT (A) and NHEK (B) cells were treated with diﬀerent concentration (0, 2, 5 and 10 ng ml1) of TGF- b1 for 24 h, and then TIMP3 expression was detected by Western blot. HaCaT (C) and NHEK (D) cells were treated with 5 ng ml1 of TGF-b1 at diﬀerent time period (0, 6, 12 and 24 h), followed by the measurement of TIMP3 expression by Western blot. *P < 0.05 or **P < 0.01 or ***P < 0.001 vs. control.

Figure Legend Snippet: Fig. 4 TGF-b1 repressed the expression of TIMP3. HaCaT (A) and NHEK (B) cells were treated with diﬀerent concentration (0, 2, 5 and 10 ng ml1) of TGF- b1 for 24 h, and then TIMP3 expression was detected by Western blot. HaCaT (C) and NHEK (D) cells were treated with 5 ng ml1 of TGF-b1 at diﬀerent time period (0, 6, 12 and 24 h), followed by the measurement of TIMP3 expression by Western blot. *P < 0.05 or **P < 0.01 or ***P < 0.001 vs. control.

Techniques Used: Expressing, Concentration Assay, Western Blot, Control

Fig. 5 TIMP3 inhibited the proliferation and migration of HaCaT cells under TGF-b1 treatment. HaCaT cells were treated with TGF-b1 (5 ng ml1) or transfected with pcDNA or pcDNA-TIMP3 prior to TGF-b1 treatment. (A) and (B) qRT-PCR for miR-132 expression in treated cells. (C) and (D) MTT assay for the proliferation ability in treated cells. (E) and (F) Transwell assay for the migration capacity in treated cells. *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + pcDNA.

Figure Legend Snippet: Fig. 5 TIMP3 inhibited the proliferation and migration of HaCaT cells under TGF-b1 treatment. HaCaT cells were treated with TGF-b1 (5 ng ml1) or transfected with pcDNA or pcDNA-TIMP3 prior to TGF-b1 treatment. (A) and (B) qRT-PCR for miR-132 expression in treated cells. (C) and (D) MTT assay for the proliferation ability in treated cells. (E) and (F) Transwell assay for the migration capacity in treated cells. *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + pcDNA.

Techniques Used: Migration, Transfection, Quantitative RT-PCR, Expressing, MTT Assay, Transwell Assay, Control

Fig. 6 MiR-132-mediated pro-proliferation and pro-migration eﬀects were antagonized by TIMP3 in TGF-b1-treated HaCaT cells. HaCaT cells were transfected with miR-132 mimics + pcDNA, miR-132 mimics + pcDNA-TIMP3, anti-miR-132 + si-NC or anti-miR-132 + si-TIMP3 prior to TGF-b1 (5 ng ml1) treatment, followed by the detection of cell proliferation ability by MTT assay (A) and (B), cell migration capacity by transwell assay (C) and (D). *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + miR-NC or TGF-b1 + miR-132 + pcDNA.

Figure Legend Snippet: Fig. 6 MiR-132-mediated pro-proliferation and pro-migration eﬀects were antagonized by TIMP3 in TGF-b1-treated HaCaT cells. HaCaT cells were transfected with miR-132 mimics + pcDNA, miR-132 mimics + pcDNA-TIMP3, anti-miR-132 + si-NC or anti-miR-132 + si-TIMP3 prior to TGF-b1 (5 ng ml1) treatment, followed by the detection of cell proliferation ability by MTT assay (A) and (B), cell migration capacity by transwell assay (C) and (D). *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + miR-NC or TGF-b1 + miR-132 + pcDNA.

Techniques Used: Migration, Transfection, MTT Assay, Transwell Assay, Control

Incubation:

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3
Article Snippet: Equal amounts of protein lysates (100 μg) was subjected to 10% SDS-PAGE and then electrotransferred to PVDF membrances (Bio-Rad, Hercules, CA, USA). .. Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specific primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or β-actin (sc-517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 °C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000). .. The protein bands were visualized by the ECL assay (GE Healthcare, Princeton, NJ, USA).

other:

Article Title: Tissue Inhibitor of Metalloproteinase 1 Influences Vascular Adaptations to Chronic Alterations in Blood Flow.
Article Snippet: Remodeling of the skeletal muscle microvasculature involves the coordinated actions of matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitor of metalloproteinases (TIMPs).. We hypothesized that the loss of TIMP1 would enhance both ischemia and flow-induced vascular remodeling by increasing MMP activity.. TIMP1 deficient (Timp1 / ) and wild-type (WT) C57BL/6 mice underwent unilateral femoral artery (FA) ligation or were treated with prazosin, an alpha-1 adrenergic receptor antagonist, in order to investigate vascular remodeling to altered flow.

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Journal: RSC advances

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3.

doi: 10.1039/c8ra10552a

Figure Lengend Snippet: Fig. 3 TIMP3 was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 30-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. **P < 0.01 or ***P < 0.001 vs. respective control.

Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specic primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or b-actin (sc517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

Techniques: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Construct, Transfection, Quantitative RT-PCR, Western Blot, Control

Journal: RSC advances

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3.

doi: 10.1039/c8ra10552a

Figure Lengend Snippet: Fig. 4 TGF-b1 repressed the expression of TIMP3. HaCaT (A) and NHEK (B) cells were treated with diﬀerent concentration (0, 2, 5 and 10 ng ml1) of TGF- b1 for 24 h, and then TIMP3 expression was detected by Western blot. HaCaT (C) and NHEK (D) cells were treated with 5 ng ml1 of TGF-b1 at diﬀerent time period (0, 6, 12 and 24 h), followed by the measurement of TIMP3 expression by Western blot. *P < 0.05 or **P < 0.01 or ***P < 0.001 vs. control.

Techniques: Expressing, Concentration Assay, Western Blot, Control

Journal: RSC advances

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3.

doi: 10.1039/c8ra10552a

Figure Lengend Snippet: Fig. 5 TIMP3 inhibited the proliferation and migration of HaCaT cells under TGF-b1 treatment. HaCaT cells were treated with TGF-b1 (5 ng ml1) or transfected with pcDNA or pcDNA-TIMP3 prior to TGF-b1 treatment. (A) and (B) qRT-PCR for miR-132 expression in treated cells. (C) and (D) MTT assay for the proliferation ability in treated cells. (E) and (F) Transwell assay for the migration capacity in treated cells. *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + pcDNA.

Techniques: Migration, Transfection, Quantitative RT-PCR, Expressing, MTT Assay, Transwell Assay, Control

Journal: RSC advances

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3.

doi: 10.1039/c8ra10552a

Figure Lengend Snippet: Fig. 6 MiR-132-mediated pro-proliferation and pro-migration eﬀects were antagonized by TIMP3 in TGF-b1-treated HaCaT cells. HaCaT cells were transfected with miR-132 mimics + pcDNA, miR-132 mimics + pcDNA-TIMP3, anti-miR-132 + si-NC or anti-miR-132 + si-TIMP3 prior to TGF-b1 (5 ng ml1) treatment, followed by the detection of cell proliferation ability by MTT assay (A) and (B), cell migration capacity by transwell assay (C) and (D). *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + miR-NC or TGF-b1 + miR-132 + pcDNA.

Techniques: Migration, Transfection, MTT Assay, Transwell Assay, Control

TIMP3 was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 3′-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. ** P < 0.01 or *** P < 0.001 vs. respective control.

Journal: RSC Advances

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3

doi: 10.1039/c8ra10552a

Figure Lengend Snippet: TIMP3 was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 3′-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. ** P < 0.01 or *** P < 0.001 vs. respective control.

Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specific primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or β-actin (sc-517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 °C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

Techniques: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Construct, Transfection, Quantitative RT-PCR, Western Blot, Control

TGF-β1 repressed the expression of TIMP3. HaCaT (A) and NHEK (B) cells were treated with different concentration (0, 2, 5 and 10 ng ml −1 ) of TGF-β1 for 24 h, and then TIMP3 expression was detected by Western blot. HaCaT (C) and NHEK (D) cells were treated with 5 ng ml −1 of TGF-β1 at different time period (0, 6, 12 and 24 h), followed by the measurement of TIMP3 expression by Western blot. * P < 0.05 or ** P < 0.01 or *** P < 0.001 vs. control.

Journal: RSC Advances

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3

doi: 10.1039/c8ra10552a

Figure Lengend Snippet: TGF-β1 repressed the expression of TIMP3. HaCaT (A) and NHEK (B) cells were treated with different concentration (0, 2, 5 and 10 ng ml −1 ) of TGF-β1 for 24 h, and then TIMP3 expression was detected by Western blot. HaCaT (C) and NHEK (D) cells were treated with 5 ng ml −1 of TGF-β1 at different time period (0, 6, 12 and 24 h), followed by the measurement of TIMP3 expression by Western blot. * P < 0.05 or ** P < 0.01 or *** P < 0.001 vs. control.

Techniques: Expressing, Concentration Assay, Western Blot, Control

TIMP3 inhibited the proliferation and migration of HaCaT cells under TGF-β1 treatment. HaCaT cells were treated with TGF-β1 (5 ng ml −1 ) or transfected with pcDNA or pcDNA-TIMP3 prior to TGF-β1 treatment. (A) and (B) qRT-PCR for miR-132 expression in treated cells. (C) and (D) MTT assay for the proliferation ability in treated cells. (E) and (F) Transwell assay for the migration capacity in treated cells. * P < 0.05 or ** P < 0.01 vs. control or TGF-β1 + pcDNA.

Journal: RSC Advances

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3

doi: 10.1039/c8ra10552a

Figure Lengend Snippet: TIMP3 inhibited the proliferation and migration of HaCaT cells under TGF-β1 treatment. HaCaT cells were treated with TGF-β1 (5 ng ml −1 ) or transfected with pcDNA or pcDNA-TIMP3 prior to TGF-β1 treatment. (A) and (B) qRT-PCR for miR-132 expression in treated cells. (C) and (D) MTT assay for the proliferation ability in treated cells. (E) and (F) Transwell assay for the migration capacity in treated cells. * P < 0.05 or ** P < 0.01 vs. control or TGF-β1 + pcDNA.

Techniques: Migration, Transfection, Quantitative RT-PCR, Expressing, MTT Assay, Transwell Assay, Control

MiR-132-mediated pro-proliferation and pro-migration effects were antagonized by TIMP3 in TGF-β1-treated HaCaT cells. HaCaT cells were transfected with miR-132 mimics + pcDNA, miR-132 mimics + pcDNA-TIMP3, anti-miR-132 + si-NC or anti-miR-132 + si-TIMP3 prior to TGF-β1 (5 ng ml −1 ) treatment, followed by the detection of cell proliferation ability by MTT assay (A) and (B), cell migration capacity by transwell assay (C) and (D). * P < 0.05 or ** P < 0.01 vs. control or TGF-β1 + miR-NC or TGF-β1 + miR-132 + pcDNA.

Journal: RSC Advances

Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3

doi: 10.1039/c8ra10552a

Figure Lengend Snippet: MiR-132-mediated pro-proliferation and pro-migration effects were antagonized by TIMP3 in TGF-β1-treated HaCaT cells. HaCaT cells were transfected with miR-132 mimics + pcDNA, miR-132 mimics + pcDNA-TIMP3, anti-miR-132 + si-NC or anti-miR-132 + si-TIMP3 prior to TGF-β1 (5 ng ml −1 ) treatment, followed by the detection of cell proliferation ability by MTT assay (A) and (B), cell migration capacity by transwell assay (C) and (D). * P < 0.05 or ** P < 0.01 vs. control or TGF-β1 + miR-NC or TGF-β1 + miR-132 + pcDNA.

Techniques: Migration, Transfection, MTT Assay, Transwell Assay, Control

Levels of testicular TIMPs and MPs in response to MEHP exposure by Western blot analysis. Whole-testis homogenates from 28-day-old C57BL/6J mice and total protein from primary Sertoli cell-germ cell cocultures exposed to MEHP (1 g/kg) are examined, and the quantified relative protein levels are shown as column charts. TIMP1, TIMP2, TIMP3, and TIMP4 protein levels (A) and MMP2, MMP9, ADAM10, and ADAM17 protein levels (B) are determined in vivo. After MEHP exposure, TIMP1 and TIMP2 levels are decreased, and MMP2 levels are increased for a short period. C) MMP2 and TIMP2 protein levels in primary coculture cells are analyzed. TIMP2 expression is observed only in primary Sertoli cells and is significantly decreased after MEHP (200 μM) exposure. GC, germ cells; SC, Sertoli cells. ACTB served as the loading control. Asterisks denote significant differences between the treatment and the control (* P < 0.05, Student t-test).

Journal: Biology of Reproduction

Article Title: TNF Alpha-Mediated Disruption of Spermatogenesis in Response to Sertoli Cell Injury in Rodents Is Partially Regulated by MMP2 1

doi: 10.1095/biolreprod.108.073122

Figure Lengend Snippet: Levels of testicular TIMPs and MPs in response to MEHP exposure by Western blot analysis. Whole-testis homogenates from 28-day-old C57BL/6J mice and total protein from primary Sertoli cell-germ cell cocultures exposed to MEHP (1 g/kg) are examined, and the quantified relative protein levels are shown as column charts. TIMP1, TIMP2, TIMP3, and TIMP4 protein levels (A) and MMP2, MMP9, ADAM10, and ADAM17 protein levels (B) are determined in vivo. After MEHP exposure, TIMP1 and TIMP2 levels are decreased, and MMP2 levels are increased for a short period. C) MMP2 and TIMP2 protein levels in primary coculture cells are analyzed. TIMP2 expression is observed only in primary Sertoli cells and is significantly decreased after MEHP (200 μM) exposure. GC, germ cells; SC, Sertoli cells. ACTB served as the loading control. Asterisks denote significant differences between the treatment and the control (* P < 0.05, Student t-test).

Article Snippet: Total cellular proteins (30 μg) were tested using primary antibodies against TIMP1, TIMP2, TIMP3, and TIMP4 (1:1000; Chemicon, Temecula, CA); matrix MP 2 (MMP2) and MMP9 (1:1000; Abcam Inc., Cambridge, MA); FASL (1:500; Santa Cruz Biotechnology Inc., Santa Cruz, CA); TNFA (1:500; R&D Systems Inc., Minneapolis, MN); a distintegrin and MP 10 (ADAM10) (1:500; R&D Systems Inc.), ADAM17 (1:500; Abcam Inc.); and β-actin (ACTB) (1:500; Santa Cruz Biotechnology Inc.) coupled with horseradish peroxidase-conjugated secondary antibodies (1:5000; Santa Cruz Biotechnology Inc.).

Techniques: Western Blot, In Vivo, Expressing

A proposed working model of the complex Sertoli cell-germ cell interaction describing the mechanism by which Sertoli cells trigger germ cell apoptosis following MEHP-induced Sertoli cell injury. First, MEHP-induced decreases in TIMP2 expression in Sertoli cells allow for the increased activation of MMP2. Active MMP2 can then cleave membrane-bound TNFA produced by germ cells to generate the soluble active form of TNFA. The binding of sTNFA with Sertoli cell TNFRSF1A triggers the NFKB1 signaling pathway and robustly induces FASL expression. An increase in Sertoli cell FASL allows for the instigation of germ cell apoptosis by the classic death receptor pathway.

Journal: Biology of Reproduction

Article Title: TNF Alpha-Mediated Disruption of Spermatogenesis in Response to Sertoli Cell Injury in Rodents Is Partially Regulated by MMP2 1

doi: 10.1095/biolreprod.108.073122

Figure Lengend Snippet: A proposed working model of the complex Sertoli cell-germ cell interaction describing the mechanism by which Sertoli cells trigger germ cell apoptosis following MEHP-induced Sertoli cell injury. First, MEHP-induced decreases in TIMP2 expression in Sertoli cells allow for the increased activation of MMP2. Active MMP2 can then cleave membrane-bound TNFA produced by germ cells to generate the soluble active form of TNFA. The binding of sTNFA with Sertoli cell TNFRSF1A triggers the NFKB1 signaling pathway and robustly induces FASL expression. An increase in Sertoli cell FASL allows for the instigation of germ cell apoptosis by the classic death receptor pathway.

Techniques: Expressing, Activation Assay, Produced, Binding Assay

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specic primary antibodies against timp3 (Santa Cruz Biotechnology)

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