ϐ actin  (Millipore)


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  • 99
    Name:
    Anti beta Actin antibody
    Description:
    The ACTB β actin gene is mapped to human chromosome 7p22 1 β Actin is the most abundant protein localized to the cytoplasm ACTB is expressed ubiquitously Actin is one of the most conserved eukaryotic proteins it is expressed in mammals and birds Four of the actin isoforms represent the differentiation markers of muscle tissues and two are found in almost all cells Anti β Actin antibody Mouse Monoclonal mouse IgG1 isotype is derived from the AC 15 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from BALB c mice immunized with a slightly modified synthetic b cytoplasmic actin N terminal peptide Ac Asp Asp Asp Ile Ala Ala Leu Val Ile Asp Asn Gly Ser Gly Lys conjugated to KLH In staining of chicken gizzard ultrathin tissue cryosections the antibody labels the dense bodies and longitudinal channels linking consecutive dense bodies that are also occupied by desmin and the membrane associated dense plaque It does not stain adult cardiac and skeletal muscles except for traces due to contaminations of the sample with non muscle cells or if embryonic tissue is being used
    Catalog Number:
    a1978
    Price:
    None
    Applications:
    The actin in cells of various species and tissue origin is very similar in their immunological and physical properties. As a consequence, it has been difficult to produce potent antisera to this protein. Therefore the availability of monoclonal antibodies to β-actin provides a specific and useful tool in studying the intracellular distribution of β-actin and the static and dynamic aspects of the cytoskeleton.Monoclonal anti-beta-actin antibody can be used for microarray, indirect immunofluorescence, and immunohistochemical analyses. Furthermore, the product has been used for immunocytochemistry at 10-40 μg/mL using human foreskin fibroblasts. The antibody has also been used for western blot at 0.5-1 μg/mL using cell extract of human foreskin fibroblasts or chicken fibroblasts.The antibody can be used for staining of acetone-fixed frozen sections, EM preparations, and microinjection experiments. B5, ethanol, methacam, or Bouin′s solutions can be used as fixatives. The epitope recognized by the antibody is resistant to formalin-fixation and paraffin-embedding.It has been used in Immunoblot analysis.
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    Structured Review

    Millipore ϐ actin
    Anti beta Actin antibody
    The ACTB β actin gene is mapped to human chromosome 7p22 1 β Actin is the most abundant protein localized to the cytoplasm ACTB is expressed ubiquitously Actin is one of the most conserved eukaryotic proteins it is expressed in mammals and birds Four of the actin isoforms represent the differentiation markers of muscle tissues and two are found in almost all cells Anti β Actin antibody Mouse Monoclonal mouse IgG1 isotype is derived from the AC 15 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from BALB c mice immunized with a slightly modified synthetic b cytoplasmic actin N terminal peptide Ac Asp Asp Asp Ile Ala Ala Leu Val Ile Asp Asn Gly Ser Gly Lys conjugated to KLH In staining of chicken gizzard ultrathin tissue cryosections the antibody labels the dense bodies and longitudinal channels linking consecutive dense bodies that are also occupied by desmin and the membrane associated dense plaque It does not stain adult cardiac and skeletal muscles except for traces due to contaminations of the sample with non muscle cells or if embryonic tissue is being used
    https://www.bioz.com/result/ϐ actin/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ϐ actin - by Bioz Stars, 2020-07
    99/100 stars

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    Related Articles

    Blocking Assay:

    Article Title: Modeling the natural history of Pelizaeus–Merzbacher disease
    Article Snippet: .. After a dry transfer and blocking, membranes were incubated with antibodies as follows: mouse anti-β-actin (1:8000, Sigma), rabbit anti-PLP (1:10,000, gift from Dr. 1. .. Griffiths), and rabbit anti-MBP (1:1000, Millipore).

    FLAG-tag:

    Article Title: Seven in Absentia Homolog 2 (Siah2) Protein Is a Regulator of NF-E2-related Factor 2 (Nrf2) *Seven in Absentia Homolog 2 (Siah2) Protein Is a Regulator of NF-E2-related Factor 2 (Nrf2) * ♦
    Article Snippet: .. Anti-c-Myc, anti-DYKDDDDK (FLAG) tag, and anti-HA monoclonal antibodies were purchased from Wako; anti-phosphoserine antibody was from BD Biosciences; anti-human ubiquitin antibody was from Enzo Life Sciences (Farmingdale, NY); anti-human Siah2 antibody was from Santa Cruz Biotechnology (Santa Cruz, CA); anti-human β-actin antibody was from Sigma; and horseradish peroxidase-conjugated anti-rabbit and anti-mouse IgG antibodies were from Bio-Rad. ..

    Incubation:

    Article Title: SPARCL1 suppresses metastasis in prostate cancer
    Article Snippet: .. Membranes were incubated with chicken polyclonal antibody specific for human SPARCL1 (Abcam, Cambridge, United Kingdom) at a 1:2000 dilution, and then the membranes were re‐probed using mouse monoclonal antibody specific for human anti‐β‐actin (Sigma Chemical Company, Saint Louis, MO, USA). .. The human gene SPARCL1 ORF (RC207583, OriGene Technologies, Inc, Rockville, MD) was subcloned into pBMN‐I‐GFP (Addgene Inc., Cambridge, MA, USA) to obtain a pBMN‐SPARCL1‐I‐GFP plasmid.

    Article Title: The translocator protein ligand [18F]DPA-714 images glioma and activated microglia in vivo
    Article Snippet: .. Incubation with the primary antibodies diluted in PBST with 3% of BSA for the anti-TSPO antibody and 3% non-fat dry milk for the anti-β-actin antibody, respectively (1/10,000 dilution of the anti-rat TSPO antibody NP155 [ ] generously provided by Dr. M. Higuchi, NIRS, Japan, and 1/5,000 dilution of the anti-β-actin antibody purchased from Sigma Aldrich), was performed overnight at 4°C. .. After three wash steps, the membrane was incubated with the appropriate secondary antibody (1/2,000 dilution) in 1% of BSA or 1% non-fat dry milk in PBST for 1 h at room temperature.

    Article Title: Modeling the natural history of Pelizaeus–Merzbacher disease
    Article Snippet: .. After a dry transfer and blocking, membranes were incubated with antibodies as follows: mouse anti-β-actin (1:8000, Sigma), rabbit anti-PLP (1:10,000, gift from Dr. 1. .. Griffiths), and rabbit anti-MBP (1:1000, Millipore).

    Article Title: Photodynamic therapy inhibits p-glycoprotein mediated multidrug resistance via JNK activation in human hepatocellular carcinoma using the photosensitizer pheophorbide a
    Article Snippet: .. The membrane was blocked with 10% non-fat milk in Tris buffered saline containing Tween-20 (TBS-T) (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, 0.1% Tween-20) and then incubated with primary human antibodies against β-actin (Sigma), Bcl-2, JNK, p-JNK, p-ERK, procaspase-3 (Santa Cruz), caspase-9 (Stressgen), and P-glycoprotein (Merck) in TBS-T. After incubation with the secondary antibody conjugated with horseradish peroxidase, immunodetected proteins were visualized by using an enhanced chemiluminescence assay kit (Amersham Life Science). ..

    Article Title: ORP150/HSP12A Regulates Purkinje Cell Survival: A Role for Endoplasmic Reticulum Stress in Cerebellar Development
    Article Snippet: .. Tissue extracts were prepared in PBS containing NP-40 (1%), and proteins were separated by SDS-PAGE and transferred to polyvinylidene difluoride paper, followed by incubation with either anti-human ORP150 antibody (1 μg/ml) or anti-human β-actin antibody (1000× dilution; Sigma), the latter as an internal control. .. Where indicated, Western blotting using anti-KDEL monoclonal antibody (Stressgen Biotechnologies Co.; 0.2 μg/ml) was used to assess levels of GRP78 (78 kDa glucose-regulated protein; ).

    Chemiluminescence Immunoassay:

    Article Title: Photodynamic therapy inhibits p-glycoprotein mediated multidrug resistance via JNK activation in human hepatocellular carcinoma using the photosensitizer pheophorbide a
    Article Snippet: .. The membrane was blocked with 10% non-fat milk in Tris buffered saline containing Tween-20 (TBS-T) (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, 0.1% Tween-20) and then incubated with primary human antibodies against β-actin (Sigma), Bcl-2, JNK, p-JNK, p-ERK, procaspase-3 (Santa Cruz), caspase-9 (Stressgen), and P-glycoprotein (Merck) in TBS-T. After incubation with the secondary antibody conjugated with horseradish peroxidase, immunodetected proteins were visualized by using an enhanced chemiluminescence assay kit (Amersham Life Science). ..

    Western Blot:

    Article Title: FAK activation is required for IGF1R-mediated regulation of EMT, migration, and invasion in mesenchymal triple negative breast cancer cells
    Article Snippet: .. ZEB-1 antibody was from Santa Cruz Biotechnology, Inc. Antibodies against β-actin, and vimentin purchased from Sigma (Saint Louis, MO, USA) and pFAK (Tyr397), total FAK, and E-cadherin from BD Biosciences were used for Western blots according to standard protocol. .. Bound primary antibodies were detected with peroxidase-coupled secondary antibodies (Southern BioTech; Birmingham, AL, USA) and developed by enhanced chemiluminescence (Luminata Classico Western HRP substrate; EMD Millipore Corp.; Billerica, MA, USA).

    SDS Page:

    Article Title: ORP150/HSP12A Regulates Purkinje Cell Survival: A Role for Endoplasmic Reticulum Stress in Cerebellar Development
    Article Snippet: .. Tissue extracts were prepared in PBS containing NP-40 (1%), and proteins were separated by SDS-PAGE and transferred to polyvinylidene difluoride paper, followed by incubation with either anti-human ORP150 antibody (1 μg/ml) or anti-human β-actin antibody (1000× dilution; Sigma), the latter as an internal control. .. Where indicated, Western blotting using anti-KDEL monoclonal antibody (Stressgen Biotechnologies Co.; 0.2 μg/ml) was used to assess levels of GRP78 (78 kDa glucose-regulated protein; ).

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  • 95
    Millipore β actin
    Effects of repeated TM stimulation on fibroblasts’ morphology and GRP78/BiP expression. Primary cultured fibroblasts were treated with 1μg/ml TM or DMSO for 5 minutes per day 3 days in series. After this repeated TM or DMSO stimulation, medium was changed to DMEM with 2% horse serum (Basal medium condition) and incubated for 12h to induce differentiation. Primary cultured fibroblasts were treated with 1μg/ml TM or DMSO for 5 minutes per day 3 days in series. (a) Just after this repeated TM or DMSO stimulation, the cells were observed (upper panels) and stained by anti-Bip antibody (bottom panels). (b) The cells treated with TM (Rep-TM) or DMSO (Rep-DM/Rep-DMSO) cultured in the culture condition medium (C.C.) or in the Basal medium condition for differentiation (M.C.) were collected and lysed. Western blot analysis was performed using an anti-Bip or <t>anti-β-actin</t> primary antibody (upper panels). Quantitative data were obtained by densitometry of the bands. Data are expressed as the mean ± SEM for at least three independent experiments (shown as a ratio of the Rep-DM C.C.). The P value was compared with the control and calculated by Student's T test. (c) Left and middle panels show the cells treated with TM (Rep-TM) or DMSO (Rep-DMSO) cultured at Basal medium condition. Right panel shows the cells treated with TGF-β1 after the incubation at the basal medium condition.
    β Actin, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 2095 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β actin/product/Millipore
    Average 95 stars, based on 2095 article reviews
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    92
    Millipore filamin a
    Disrupting the <t>CaR-filamin</t> interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin
    Filamin A, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 5 article reviews
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    90
    Millipore β actin c4
    Western blot analysis of hormone receptors. Cell lysates from BT474 were analyzed for changes in androgen (AR), estrogen (ER) and glucocorticoid (GR) and progesterone (PGR) receptor levels. <t>β‐ACTIN</t> is used as the loading control for this
    β Actin C4, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β actin c4/product/Millipore
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    88
    Millipore ãŽâ² actin
    TGFβ inhibition augments anti–CTLA-4 immunotherapy in an autochthonous BRAF V600E PTEN -/- melanoma model. A. Mice were treated with either anti–CTLA-4 (purple, 100 μg i.p.) or IgG isotype control (black, 100 μg i.p.) every 3 days when tumors reached 60–80 mm 3 . Left : tumor volumes monitored every 3 days. Right : representative tumor photos. 6 mice/group. Representative of 3 independent experiments. B. Left : resected Braf V600E Pten -/- melanoma tissues analyzed for TGFβ1 expression by IHC and whole tissue Western blots. IHC representative of 3 tumor specimens (40x). Right : Spearman correlation between tumor volume and <t>TGFβ1/β-actin</t> density ratios from Western blots. C. Phospho-SMAD2 (pSMAD2) Western blot analysis performed following TEW-7197 treatment (25 mg/kg daily p.o. x 2 weeks). tSMAD2: total SMAD2. Representative of 2 independent experiments. D. Left : mice were treated with TEW-7197 monotherapy (25 mg/kg p.o. daily). Tumor volumes were monitored every 3 days. 6 mice/group. Representative of 3 independent experiments. Center : final tumor weights. Right : representative photos of resected tumors. ns: non-significant. E. Left : mice were treated with either IgG isotype control/vehicle control, TEW-7197(25 mg/kg p.o. daily) monotherapy, or combination anti–CTLA-4 (100 μg i.p. every 3 days)/TEW-7197. Left : tumor volumes monitored every 3 days. 6 mice/group. Black arrow, treatment initiation. Representative of 2 independent experiments. Right top : lungs resected and enumerated for metastatic foci by staggered IHC. Right bottom : Kaplan-Meier survival plot. Data analyzed by the log-rank test. F. Left : flow cytometry on tumor-infiltrating lymphocytes (TILs) at the conclusion of the experiment. 5 tumors/group. Right . All data is mean±SEM. Significance calculated using the unpaired t-test or a one-way ANOVA. * p
    ãŽâ² Actin, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ãŽâ² actin - by Bioz Stars, 2020-07
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    Image Search Results


    Effects of repeated TM stimulation on fibroblasts’ morphology and GRP78/BiP expression. Primary cultured fibroblasts were treated with 1μg/ml TM or DMSO for 5 minutes per day 3 days in series. After this repeated TM or DMSO stimulation, medium was changed to DMEM with 2% horse serum (Basal medium condition) and incubated for 12h to induce differentiation. Primary cultured fibroblasts were treated with 1μg/ml TM or DMSO for 5 minutes per day 3 days in series. (a) Just after this repeated TM or DMSO stimulation, the cells were observed (upper panels) and stained by anti-Bip antibody (bottom panels). (b) The cells treated with TM (Rep-TM) or DMSO (Rep-DM/Rep-DMSO) cultured in the culture condition medium (C.C.) or in the Basal medium condition for differentiation (M.C.) were collected and lysed. Western blot analysis was performed using an anti-Bip or anti-β-actin primary antibody (upper panels). Quantitative data were obtained by densitometry of the bands. Data are expressed as the mean ± SEM for at least three independent experiments (shown as a ratio of the Rep-DM C.C.). The P value was compared with the control and calculated by Student's T test. (c) Left and middle panels show the cells treated with TM (Rep-TM) or DMSO (Rep-DMSO) cultured at Basal medium condition. Right panel shows the cells treated with TGF-β1 after the incubation at the basal medium condition.

    Journal: PLoS ONE

    Article Title: Physiological ER Stress Mediates the Differentiation of Fibroblasts

    doi: 10.1371/journal.pone.0123578

    Figure Lengend Snippet: Effects of repeated TM stimulation on fibroblasts’ morphology and GRP78/BiP expression. Primary cultured fibroblasts were treated with 1μg/ml TM or DMSO for 5 minutes per day 3 days in series. After this repeated TM or DMSO stimulation, medium was changed to DMEM with 2% horse serum (Basal medium condition) and incubated for 12h to induce differentiation. Primary cultured fibroblasts were treated with 1μg/ml TM or DMSO for 5 minutes per day 3 days in series. (a) Just after this repeated TM or DMSO stimulation, the cells were observed (upper panels) and stained by anti-Bip antibody (bottom panels). (b) The cells treated with TM (Rep-TM) or DMSO (Rep-DM/Rep-DMSO) cultured in the culture condition medium (C.C.) or in the Basal medium condition for differentiation (M.C.) were collected and lysed. Western blot analysis was performed using an anti-Bip or anti-β-actin primary antibody (upper panels). Quantitative data were obtained by densitometry of the bands. Data are expressed as the mean ± SEM for at least three independent experiments (shown as a ratio of the Rep-DM C.C.). The P value was compared with the control and calculated by Student's T test. (c) Left and middle panels show the cells treated with TM (Rep-TM) or DMSO (Rep-DMSO) cultured at Basal medium condition. Right panel shows the cells treated with TGF-β1 after the incubation at the basal medium condition.

    Article Snippet: Equal amounts of protein were subjected to 5–20% gradient SDS-PAGE, e-PAGEL (ATTO CO., Tokyo, Japan) for GRP78/Bip or β-actin and transferred to PVDF membrane (Millipore, Bedford, MA).

    Techniques: Expressing, Cell Culture, Incubation, Staining, Western Blot

    Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin

    Journal: The Journal of investigative dermatology

    Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

    doi: 10.1038/jid.2010.414

    Figure Lengend Snippet: Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin

    Article Snippet: The following antibodies (Abs) were used in this study: monoclonal antibodies (mAbs) for α2 integrin (BD Biosciences, San Jose, CA), for human involucrin and β-actin, for c-Myc- and HA-tags (Sigma-Aldrich, St. Louis, MO), polyclonal and monoclonal Abs against E-cadherin, β-, γ-, and p120-catenin and Fyn, polyclonal Abs against Rho A, Rac1 and cdc42, and CaR, mAb against transglutaminase 1 (Santa Cruz Biotechnology, Santa Cruz, CA), mAbs for phosphotyrosine, Rho and filamin A (Millipore Corp., Temecula, CA), polyclonal Abs for human keratin 1, keratin 14 and loricrin (Covance Inc., Berkeley, CA), and rabbit anti-human CaR ( ).

    Techniques: Activation Assay

    Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane

    Journal: The Journal of investigative dermatology

    Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

    doi: 10.1038/jid.2010.414

    Figure Lengend Snippet: Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane

    Article Snippet: The following antibodies (Abs) were used in this study: monoclonal antibodies (mAbs) for α2 integrin (BD Biosciences, San Jose, CA), for human involucrin and β-actin, for c-Myc- and HA-tags (Sigma-Aldrich, St. Louis, MO), polyclonal and monoclonal Abs against E-cadherin, β-, γ-, and p120-catenin and Fyn, polyclonal Abs against Rho A, Rac1 and cdc42, and CaR, mAb against transglutaminase 1 (Santa Cruz Biotechnology, Santa Cruz, CA), mAbs for phosphotyrosine, Rho and filamin A (Millipore Corp., Temecula, CA), polyclonal Abs for human keratin 1, keratin 14 and loricrin (Covance Inc., Berkeley, CA), and rabbit anti-human CaR ( ).

    Techniques:

    Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions

    Journal: The Journal of investigative dermatology

    Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

    doi: 10.1038/jid.2010.414

    Figure Lengend Snippet: Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions

    Article Snippet: The following antibodies (Abs) were used in this study: monoclonal antibodies (mAbs) for α2 integrin (BD Biosciences, San Jose, CA), for human involucrin and β-actin, for c-Myc- and HA-tags (Sigma-Aldrich, St. Louis, MO), polyclonal and monoclonal Abs against E-cadherin, β-, γ-, and p120-catenin and Fyn, polyclonal Abs against Rho A, Rac1 and cdc42, and CaR, mAb against transglutaminase 1 (Santa Cruz Biotechnology, Santa Cruz, CA), mAbs for phosphotyrosine, Rho and filamin A (Millipore Corp., Temecula, CA), polyclonal Abs for human keratin 1, keratin 14 and loricrin (Covance Inc., Berkeley, CA), and rabbit anti-human CaR ( ).

    Techniques:

    Western blot analysis of hormone receptors. Cell lysates from BT474 were analyzed for changes in androgen (AR), estrogen (ER) and glucocorticoid (GR) and progesterone (PGR) receptor levels. β‐ACTIN is used as the loading control for this

    Journal:

    Article Title: Androgens act synergistically to enhance estrogen‐induced upregulation of human tissue kallikreins 10, 11, and 14 in breast cancer cells via a membrane bound androgen receptor), Androgens act synergistically to enhance estrogen‐induced upregulation of human tissue kallikreins 10, 11, and 14 in breast cancer cells via a membrane bound androgen receptor

    doi: 10.1016/j.molonc.2008.01.001

    Figure Lengend Snippet: Western blot analysis of hormone receptors. Cell lysates from BT474 were analyzed for changes in androgen (AR), estrogen (ER) and glucocorticoid (GR) and progesterone (PGR) receptor levels. β‐ACTIN is used as the loading control for this

    Article Snippet: The antibodies used for Western blot analysis included, AR (N‐20), ERα (D‐12), and PGR (AB‐52), and β‐ACTIN (C4) from EMD Biosciences Inc., San Diego, CA and GR (Clone 41) from BD Biosciences, San Jose, CA.

    Techniques: Western Blot

    TGFβ inhibition augments anti–CTLA-4 immunotherapy in an autochthonous BRAF V600E PTEN -/- melanoma model. A. Mice were treated with either anti–CTLA-4 (purple, 100 μg i.p.) or IgG isotype control (black, 100 μg i.p.) every 3 days when tumors reached 60–80 mm 3 . Left : tumor volumes monitored every 3 days. Right : representative tumor photos. 6 mice/group. Representative of 3 independent experiments. B. Left : resected Braf V600E Pten -/- melanoma tissues analyzed for TGFβ1 expression by IHC and whole tissue Western blots. IHC representative of 3 tumor specimens (40x). Right : Spearman correlation between tumor volume and TGFβ1/β-actin density ratios from Western blots. C. Phospho-SMAD2 (pSMAD2) Western blot analysis performed following TEW-7197 treatment (25 mg/kg daily p.o. x 2 weeks). tSMAD2: total SMAD2. Representative of 2 independent experiments. D. Left : mice were treated with TEW-7197 monotherapy (25 mg/kg p.o. daily). Tumor volumes were monitored every 3 days. 6 mice/group. Representative of 3 independent experiments. Center : final tumor weights. Right : representative photos of resected tumors. ns: non-significant. E. Left : mice were treated with either IgG isotype control/vehicle control, TEW-7197(25 mg/kg p.o. daily) monotherapy, or combination anti–CTLA-4 (100 μg i.p. every 3 days)/TEW-7197. Left : tumor volumes monitored every 3 days. 6 mice/group. Black arrow, treatment initiation. Representative of 2 independent experiments. Right top : lungs resected and enumerated for metastatic foci by staggered IHC. Right bottom : Kaplan-Meier survival plot. Data analyzed by the log-rank test. F. Left : flow cytometry on tumor-infiltrating lymphocytes (TILs) at the conclusion of the experiment. 5 tumors/group. Right . All data is mean±SEM. Significance calculated using the unpaired t-test or a one-way ANOVA. * p

    Journal: Cancer immunology research

    Article Title: Stromal Fibroblasts Mediate Anti–PD-1 Resistance via MMP-9 and Dictate TGFβ Inhibitor Sequencing in Melanoma

    doi: 10.1158/2326-6066.CIR-18-0086

    Figure Lengend Snippet: TGFβ inhibition augments anti–CTLA-4 immunotherapy in an autochthonous BRAF V600E PTEN -/- melanoma model. A. Mice were treated with either anti–CTLA-4 (purple, 100 μg i.p.) or IgG isotype control (black, 100 μg i.p.) every 3 days when tumors reached 60–80 mm 3 . Left : tumor volumes monitored every 3 days. Right : representative tumor photos. 6 mice/group. Representative of 3 independent experiments. B. Left : resected Braf V600E Pten -/- melanoma tissues analyzed for TGFβ1 expression by IHC and whole tissue Western blots. IHC representative of 3 tumor specimens (40x). Right : Spearman correlation between tumor volume and TGFβ1/β-actin density ratios from Western blots. C. Phospho-SMAD2 (pSMAD2) Western blot analysis performed following TEW-7197 treatment (25 mg/kg daily p.o. x 2 weeks). tSMAD2: total SMAD2. Representative of 2 independent experiments. D. Left : mice were treated with TEW-7197 monotherapy (25 mg/kg p.o. daily). Tumor volumes were monitored every 3 days. 6 mice/group. Representative of 3 independent experiments. Center : final tumor weights. Right : representative photos of resected tumors. ns: non-significant. E. Left : mice were treated with either IgG isotype control/vehicle control, TEW-7197(25 mg/kg p.o. daily) monotherapy, or combination anti–CTLA-4 (100 μg i.p. every 3 days)/TEW-7197. Left : tumor volumes monitored every 3 days. 6 mice/group. Black arrow, treatment initiation. Representative of 2 independent experiments. Right top : lungs resected and enumerated for metastatic foci by staggered IHC. Right bottom : Kaplan-Meier survival plot. Data analyzed by the log-rank test. F. Left : flow cytometry on tumor-infiltrating lymphocytes (TILs) at the conclusion of the experiment. 5 tumors/group. Right . All data is mean±SEM. Significance calculated using the unpaired t-test or a one-way ANOVA. * p

    Article Snippet: Primary antibodies included TGFβ (Cell Signaling, cat. number 3711), total SMAD2 (Cell signaling, cat. number 5339), phospho-SMAD2 (S465/467; Cell signaling, cat. number 3108), β-actin (Millipore, cat. number MABT1333), and MMP-9 (Millipore, cat. number MABT171) and were used at 1:1000.

    Techniques: Inhibition, Mouse Assay, Expressing, Immunohistochemistry, Western Blot, Flow Cytometry, Cytometry