polyclonal anti grb7 antibody  (Boster Bio)

Journal: Cureus

Article Title: Detecting Human Epidermal Growth Factor Receptor 2 (HER2) Amplification: Proof of Concept of an Alternative Approach

doi: 10.7759/cureus.44785

Figure Lengend Snippet: a: Section showing diffuse, strong, granular, grade 3 cytoplasmic positivity (magnification, 100x) for antibody to GRB7, b: Grade 2 cytoplasmic positivity, with a mixture of strong and weak staining patterns in almost equal proportion for PGAP3, c: Weak non-granular stain involving most of the cells (grade 1 cytoplasmic positivity) for GRB7. D. Negative (grade 0) cytoplasmic staining, with most cells showing no staining, or only weak patchy stain in a minority of the cells.

Article Snippet: Sections were cut from these blocks and then immuno-stained with GRB7 antibodies (make Elabsciences, Catalogue no. E-AB-14108) and PGAP3 antibodies (make Abbexa, Catalogue no. abx027296 and ThermoFisher, Catalogue no. PA5- 53451).

Techniques: Staining

Journal: Cureus

Article Title: Detecting Human Epidermal Growth Factor Receptor 2 (HER2) Amplification: Proof of Concept of an Alternative Approach

doi: 10.7759/cureus.44785

Figure Lengend Snippet: The relationship of the GRB7 and PGAP3 immunohistochemistry (IHC) staining with HER2 status. HER2: Human epidermal growth factor receptor 2, GRB7: growth factor receptor bound protein 7, PGAP3: post-GPI Attachment to protein phospholipase 3

Article Snippet: Sections were cut from these blocks and then immuno-stained with GRB7 antibodies (make Elabsciences, Catalogue no. E-AB-14108) and PGAP3 antibodies (make Abbexa, Catalogue no. abx027296 and ThermoFisher, Catalogue no. PA5- 53451).

Techniques: Immunohistochemistry

Journal: Cureus

Article Title: Detecting Human Epidermal Growth Factor Receptor 2 (HER2) Amplification: Proof of Concept of an Alternative Approach

doi: 10.7759/cureus.44785

Figure Lengend Snippet: The results of multivariate logistic regression with the most parsimonious fit among the different models used to predict the human epidermal growth factor receptor 2 (HER2) status. GRB7 membranous and PGAP3 cytoplasmic immunohistochemistry staining were the other competitors. GRB7: Growth factor receptor bound protein 7, PGAP3: post-GPI attachment to protein phospholipase 3

Article Snippet: Sections were cut from these blocks and then immuno-stained with GRB7 antibodies (make Elabsciences, Catalogue no. E-AB-14108) and PGAP3 antibodies (make Abbexa, Catalogue no. abx027296 and ThermoFisher, Catalogue no. PA5- 53451).

Techniques: Immunohistochemistry, Staining

Journal: Cureus

Article Title: Detecting Human Epidermal Growth Factor Receptor 2 (HER2) Amplification: Proof of Concept of an Alternative Approach

doi: 10.7759/cureus.44785

Figure Lengend Snippet: Area under the curves (AUCs) of the various immunohistochemical staining with the scores of GRB7 and PGAP3 added together, along with the sensitivity, specificity, and accuracy at the best cut-off. 95% confidence intervals are given in parentheses. GRB7: Growth factor receptor bound protein 7, PGAP3: post-GPI attachment to protein phospholipase 3

Article Snippet: Sections were cut from these blocks and then immuno-stained with GRB7 antibodies (make Elabsciences, Catalogue no. E-AB-14108) and PGAP3 antibodies (make Abbexa, Catalogue no. abx027296 and ThermoFisher, Catalogue no. PA5- 53451).

Techniques: Immunohistochemical staining, Staining

Journal: Cureus

Article Title: Detecting Human Epidermal Growth Factor Receptor 2 (HER2) Amplification: Proof of Concept of an Alternative Approach

doi: 10.7759/cureus.44785

Figure Lengend Snippet: The specificity of a negative test result in IHC HER2 2+ tumors (n=20). For a single marker, grade 0 was taken as negative. For a combination of markers, the best cut-offs as shown in Table 4 were used.

Article Snippet: Sections were cut from these blocks and then immuno-stained with GRB7 antibodies (make Elabsciences, Catalogue no. E-AB-14108) and PGAP3 antibodies (make Abbexa, Catalogue no. abx027296 and ThermoFisher, Catalogue no. PA5- 53451).

Techniques: Marker

Journal: Theranostics

Article Title: Methylation-associated silencing of miR-193a-3p promotes ovarian cancer aggressiveness by targeting GRB7 and MAPK/ERK pathways

doi: 10.7150/thno.22377

Figure Lengend Snippet: The expression of miR-193a-3p is downregulated and inversely correlates with GRB7 expression in ovarian cancers. (A) Comparison of GRB7 expression in human ovarian cancer with other tumor types using the cBioPortal database. The dots represent clinical cases, and the line in the upper and lower box plot represents the upper and lower quartile of the relative mRNA levels of all samples. (B) The Venn diagram shows miR-193a-3p and miR-193b-3p commonly predicted by miRDB, TargetScan, and Miranda. (C) A schematic diagram showing the same binding site of miR-193a-3p and miR-193b-3p at position 332-338 of human GRB7 3' UTR. (D) Western blot analysis of GRB7 expression in ovarian cancer cell lines and HOSEs. (E) qPCR analysis of miR-193a-3p ( upper ) and miR-193b-3p ( lower ) expression in ovarian cancer cell lines. SNORD48 was used as an internal control. (F) Western blot analysis showing that transient transfection of pmR-ZsGreen1-miR-193a-3p or pmR-ZsGreen1-miR-193b-3p reduced the expression of GRB7 in SKOV3 or OVCA433 cells in a dose-dependent manner. (G) A schematic diagram showing the putative miR-193a-3p binding site in wild-type and mutant GRB7 3'UTR ( Upper ). Luciferase reporter assay showing the relative luciferase activity of wild-type or mutant GRB7 3'UTR targeted by a miR-193a-3p using pmR-ZsGreen1-miR-193a-3p plasmid in HEK293 cells ( Lower ).

Article Snippet: The slides were incubated with the primary polyclonal anti-GRB7 antibody (H70, Santa Cruz Biotechnology, Inc.) at a 1:50 dilution.

Techniques: Expressing, Comparison, Binding Assay, Western Blot, Control, Transfection, Mutagenesis, Luciferase, Reporter Assay, Activity Assay, Plasmid Preparation

Journal: Theranostics

Article Title: Methylation-associated silencing of miR-193a-3p promotes ovarian cancer aggressiveness by targeting GRB7 and MAPK/ERK pathways

doi: 10.7150/thno.22377

Figure Lengend Snippet: MiR-193a-3p targets not only GRB7, but other key factors along MAPK/ERK signaling pathway in ovarian cancer cells. (A) The Venn diagram displays 77 common putative targets of miR-193a-3p predicted from the three algorithms, miRDB, miRanda and TargetScan Human 5.2, including three putative targets, ERBB4, KRAS and SOS2, involved in the MAPK/ERK signaling pathway. (B) Functional annotation analysis using the DAVID functional annotation tool ( http://david.abcc.ncifcrf.gov/ ) showing the main signaling pathways associated with these 78 common targets of miR-193a-3p . (C) Duolink proximity ligation assay (PLA) with fluorescence and confocal microscopy showing the interactions between GRB7 and SOS2 and between GRB7 and KRAS (red dots) by transient transfection of GRB7-expressing plasmid ( Upper ). Anti-Myc, anti-SOS2 and anti-KRAS were used to detect Myc-tagged GRB7 and endogenous SOS2 and KRAS. Transient transfection of HA-SOS2 also showing the interaction between SOS2 and KRAS (red dots) using PLA and anti-HA as well as anti-KRAS ( Lower ). Scale bar, 20 μm. n = 3 independent experiments. (D) Relative luciferase activity of luciferase reporters with wild-type ERBB4, SOS2 and KRAS 3'UTRs co-transfected with miR-193a-3p . (E) Western blot analysis showing that transient transfection of GFP/GRB7-expressing plasmid profoundly elevated ERK activity in HEK293, SKOV3 and OVCA433 cells. (F) Western blot analysis indicating that transient transfection of pmR-ZsGreen1-miR-193a-3p reduced the expression of GRB7, ERBB4, SOS2, KRAS and pERK1/2 in SKOV3 cells. Cell lysates of all the above cell transfectants were harvested 24 h after cell transfection.

Article Snippet: The slides were incubated with the primary polyclonal anti-GRB7 antibody (H70, Santa Cruz Biotechnology, Inc.) at a 1:50 dilution.

Techniques: Functional Assay, Protein-Protein interactions, Proximity Ligation Assay, Fluorescence, Confocal Microscopy, Transfection, Expressing, Plasmid Preparation, Luciferase, Activity Assay, Western Blot

Journal: Theranostics

Article Title: Methylation-associated silencing of miR-193a-3p promotes ovarian cancer aggressiveness by targeting GRB7 and MAPK/ERK pathways

doi: 10.7150/thno.22377

Figure Lengend Snippet: DNA methylation leads to downregulation of miR-193a-3p during the tumor development and progression of ovarian cancer cells. (A) qPCR analysis revealing a stepwise decrease in miR-193a-3p expression from Stages 1 to 4 and Grades 1 to 3 in ovarian cancers. (B) Representative photographs showing an inverse correlation between miR-193a-3p and GRB7 expression by ISH and IHC analyses for serous subtype ovarian cancer samples (early stage = Stage 1 and Grade 2, and advanced stage = Stage 2 and Grade 3) on a commercial ovarian cancer tissue array (OVC1021, Pantomics). Scale bar, 100 μm. (C) The expression of miR-193a-3p could be restored in ovarian cancer cells upon treatment with 5-Aza-dc (5 μM) for 4 days. (D) Verification of miR-193a-3p promoter methylation by MS-PCR in ovarian cancer cells upon 5-Aza-dC treatment. Treatment with 5-Aza-dC (5 μM) for 3-4 days. (E) Pyrosequencing analysis showing a progressive increase in DNA methylation with the tumor stage ( Upper ) and grade ( Lower ) of ovarian cancers.

Article Snippet: The slides were incubated with the primary polyclonal anti-GRB7 antibody (H70, Santa Cruz Biotechnology, Inc.) at a 1:50 dilution.

Techniques: DNA Methylation Assay, Expressing, Methylation

Journal: Theranostics

Article Title: Methylation-associated silencing of miR-193a-3p promotes ovarian cancer aggressiveness by targeting GRB7 and MAPK/ERK pathways

doi: 10.7150/thno.22377

Figure Lengend Snippet: Clinico-pathological correlation of the expression of miR193a-3p in an ovarian cancer tissue array (OVC1021). The 4-fold and 3-fold cut-off points of miR-193a-3p and GRB7, respectively, were determined by ROC analysis.

Article Snippet: The slides were incubated with the primary polyclonal anti-GRB7 antibody (H70, Santa Cruz Biotechnology, Inc.) at a 1:50 dilution.

Techniques: Expressing

Journal: Theranostics

Article Title: Methylation-associated silencing of miR-193a-3p promotes ovarian cancer aggressiveness by targeting GRB7 and MAPK/ERK pathways

doi: 10.7150/thno.22377

Figure Lengend Snippet: Re-introduction of miR-193a-3p suppresses cell proliferation, cell migration and invasion capacities of ovarian cancer cells. (A) qPCR and Western blot analyses of miR193a-3p and GRB7 expression in stable miR-193a-3p-expressing SKOV3 and OVCA433 cells. SNORD48 was used as an internal control. (B) XTT proliferation assay, (C) Transwell cell migration and invasion assays showing that stable re-expression of miR-193a-3p inhibited cell proliferation rates, cell migration and invasion abilities of SKOV3 and OVCA433 ovarian cancer cells.

Article Snippet: The slides were incubated with the primary polyclonal anti-GRB7 antibody (H70, Santa Cruz Biotechnology, Inc.) at a 1:50 dilution.

Techniques: Migration, Western Blot, Expressing, Control, Proliferation Assay

Journal: Theranostics

Article Title: Methylation-associated silencing of miR-193a-3p promotes ovarian cancer aggressiveness by targeting GRB7 and MAPK/ERK pathways

doi: 10.7150/thno.22377

Figure Lengend Snippet: The functional role of GRB7 knockdown on cell proliferation, migration, invasion, and in vivo tumorigenicity. (A) Western blot analysis showing the levels of GRB7 knockdown (KD) and scrambled control (SCR) in SKOV3 cells. (B) XTT proliferation assay and (C) Transwell cell migration and invasion assays showing that knockdown of GRB7 inhibited cell proliferation, migration and invasion. (D) The 3-4-week-old female nude mice were i.p . injected with GRB7 knockdown or scrambled control SKOV3 cells and photographed on Day 60. Yellow arrows indicate the location of tumor nodules. The number of tumor nodules on the last day in each group are illustrated by bar charts (N=5).

Article Snippet: The slides were incubated with the primary polyclonal anti-GRB7 antibody (H70, Santa Cruz Biotechnology, Inc.) at a 1:50 dilution.

Techniques: Functional Assay, Knockdown, Migration, In Vivo, Western Blot, Control, Proliferation Assay, Injection

Journal: Theranostics

Article Title: Methylation-associated silencing of miR-193a-3p promotes ovarian cancer aggressiveness by targeting GRB7 and MAPK/ERK pathways

doi: 10.7150/thno.22377

Figure Lengend Snippet: DOX-induced miR-193a-3p inhibits cell migration and invasion in vitro and results in complete inhibition of tumor growth. (A) qPCR and (B) Western blot analysis showing induction of the expression of GRB7 by 1 μg/ml doxycycline (Dox) in a time-dependent manner. SNORD48 was used as an internal control. (C) XTT cell proliferation assay showing the effect of inducible miR-193a-3p on SKOV3 cells. (D) Transwell cell migration/invasion assays demonstrating the suppressive effect of inducible miR-193a-3p on the migration and invasion capacities of SKOV3 ovarian cancer cells. The induction of miR-193a-3p for cell proliferation and cell migration/invasion using 1 μg/ml Dox. (E) Schematic overview of miR-193a-3p expression in mice and its effect on the suppression of ovarian tumor growth in the intraperitoneal cavity of nude mice. (F) The 3-4-week-old female mice received an intraperitoneal injection (i.p.) of 1x10 6 miR-193a-3p -inducible SKOV3 cells. When palpable tumors formed on Day 20, all mice were treated with or without 2 mg/mL Dox in the drinking water until they were sacrificed on Day 60. Representative photographs of tumor nodules collected after induction of miR-193a-3p on Day 60. The table shows that only 1 out of 4 mice had tumor nodules in the Dox-inducible miR-193a-3p group, while all mice (4 out of 4) developed tumor nodules in the control group. The number of tumor nodules on the last day is illustrated for each group by bar charts (N=4).

Article Snippet: The slides were incubated with the primary polyclonal anti-GRB7 antibody (H70, Santa Cruz Biotechnology, Inc.) at a 1:50 dilution.

Techniques: Migration, In Vitro, Inhibition, Western Blot, Expressing, Control, Proliferation Assay, Injection