Name: mouse monoclonal elafin
Brand: Hycult Biotech
Rating: 92 (9 reviews)

mouse monoclonal elafin (Hycult Biotech)

Hycult Biotech mouse monoclonal elafin

76NE6, 76NF2V, and 76NE7 cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral <t>elafin</t> (E). (A) Cells were harvested 48 hrs post-treatment and subjected to western blot analysis with <t>indicated</t> <t>antibodies.</t> 76NE6 cells cultured in D3-media (no growth factors) were prepared as a control (C) for elafin expression. Relative elafin levels were calculated by densitometry, displayed above. (B) Growth was monitored by trypan blue exclusion test every 24 hours for 96 hours. S . 76NE7 cells overexpressing elafin showed a statistically significant difference (t-test), in cell number at 72 hours (vs. PBS p=0.0122 and vs. Ad-luc p=0.0125) and 96 hours (vs. PBS p<0.0001 and vs. Ad-luc p<0.0001). (C) Proliferation was assessed 48 hours post-treatment by measuring BrdU incorporation; repeated in triplicate. (D) Apoptosis was assessed 72 hours post-treatment by TUNEL assay. All experiments were repeated in triplicate.

Mouse Monoclonal Elafin, supplied by Hycult Biotech, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse monoclonal elafin - by Bioz Stars, 2025-11

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76NE6, 76NF2V, and 76NE7 cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). (A) Cells were harvested 48 hrs post-treatment and subjected to western blot analysis with indicated antibodies. 76NE6 cells cultured in D3-media (no growth factors) were prepared as a control (C) for elafin expression. Relative elafin levels were calculated by densitometry, displayed above. (B) Growth was monitored by trypan blue exclusion test every 24 hours for 96 hours. S . 76NE7 cells overexpressing elafin showed a statistically significant difference (t-test), in cell number at 72 hours (vs. PBS p=0.0122 and vs. Ad-luc p=0.0125) and 96 hours (vs. PBS p<0.0001 and vs. Ad-luc p<0.0001). (C) Proliferation was assessed 48 hours post-treatment by measuring BrdU incorporation; repeated in triplicate. (D) Apoptosis was assessed 72 hours post-treatment by TUNEL assay. All experiments were repeated in triplicate.

Journal:

Article Title: The Neutrophil Elastase Inhibitor, Elafin, Triggers Rb-Mediated Growth Arrest and Caspase-Dependent Apoptosis in Breast Cancer

doi: 10.1158/0008-5472.CAN-10-1547

Figure Lengend Snippet: 76NE6, 76NF2V, and 76NE7 cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). (A) Cells were harvested 48 hrs post-treatment and subjected to western blot analysis with indicated antibodies. 76NE6 cells cultured in D3-media (no growth factors) were prepared as a control (C) for elafin expression. Relative elafin levels were calculated by densitometry, displayed above. (B) Growth was monitored by trypan blue exclusion test every 24 hours for 96 hours. S . 76NE7 cells overexpressing elafin showed a statistically significant difference (t-test), in cell number at 72 hours (vs. PBS p=0.0122 and vs. Ad-luc p=0.0125) and 96 hours (vs. PBS p<0.0001 and vs. Ad-luc p<0.0001). (C) Proliferation was assessed 48 hours post-treatment by measuring BrdU incorporation; repeated in triplicate. (D) Apoptosis was assessed 72 hours post-treatment by TUNEL assay. All experiments were repeated in triplicate.

Article Snippet: Primary antibodies used were mouse monoclonal elafin (HM2063; HyCult Biotechnology), mouse monoclonal Rb 4H1 (Cell Signaling Technology), rabbit polyclonal phospho-Rb serine 807/811 (Cell Signaling Technology), rabbit polyclonal phospho-Rb serine 780 (Cell Signaling Techonology), rabbit polyclonal PARP (Cell Signaling Technology), rabbit polyclonal Caspase 3 (Cell Signaling Technology), rabbit polyclonal Cleaved Caspase 3 Asp 175 (Cell Signaling Technology), and mouse monoclonal Actin (Chemicon International, Inc.).

Techniques: Luciferase, Western Blot, Cell Culture, Expressing, BrdU Incorporation Assay, TUNEL Assay

MCF-7, ZR75.1, T47D, MDA-MB-157, MDA-MB-436, and MDA-MB-468 cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). (A) Cells were harvested 48 hrs post-treatment and subjected to western blot analysis with indicated antibodies. (B) Growth was monitored by trypan blue exclusion test every 24 hours for 96 hours. * Elafin expression caused a statistically significant difference in cell number. (C) Proliferation was assessed 48 hours post-treatment by measuring BrdU incorporation (D) Apoptosis was assessed 72 hours post-treatment by TUNEL assay. All experiments were repeated in triplicate.

Journal:

Article Title: The Neutrophil Elastase Inhibitor, Elafin, Triggers Rb-Mediated Growth Arrest and Caspase-Dependent Apoptosis in Breast Cancer

doi: 10.1158/0008-5472.CAN-10-1547

Figure Lengend Snippet: MCF-7, ZR75.1, T47D, MDA-MB-157, MDA-MB-436, and MDA-MB-468 cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). (A) Cells were harvested 48 hrs post-treatment and subjected to western blot analysis with indicated antibodies. (B) Growth was monitored by trypan blue exclusion test every 24 hours for 96 hours. * Elafin expression caused a statistically significant difference in cell number. (C) Proliferation was assessed 48 hours post-treatment by measuring BrdU incorporation (D) Apoptosis was assessed 72 hours post-treatment by TUNEL assay. All experiments were repeated in triplicate.

Techniques: Luciferase, Western Blot, Expressing, BrdU Incorporation Assay, TUNEL Assay

(A) MDA-MB-468 were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E) for indicated times, lysates were subjected to western blot analysis with indicated antibodies. (B) Treated cells were simultaneously incubated with PBS, DMSO, or 50 μM of zVad-fmk; viability was measure by MTT assay every 24 hours for 120 hours. Eight replicates from two independent experiments were compiled and expressed as a percentage of the PBS control. Statistical significance was calculated by the t-test for no treatment vs. elafin-alone (1), luciferase alone vs. elafin alone (2), and elafin + DMSO vs. elafin + zVad-fmk are displayed as a table.

Journal:

Article Title: The Neutrophil Elastase Inhibitor, Elafin, Triggers Rb-Mediated Growth Arrest and Caspase-Dependent Apoptosis in Breast Cancer

doi: 10.1158/0008-5472.CAN-10-1547

Figure Lengend Snippet: (A) MDA-MB-468 were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E) for indicated times, lysates were subjected to western blot analysis with indicated antibodies. (B) Treated cells were simultaneously incubated with PBS, DMSO, or 50 μM of zVad-fmk; viability was measure by MTT assay every 24 hours for 120 hours. Eight replicates from two independent experiments were compiled and expressed as a percentage of the PBS control. Statistical significance was calculated by the t-test for no treatment vs. elafin-alone (1), luciferase alone vs. elafin alone (2), and elafin + DMSO vs. elafin + zVad-fmk are displayed as a table.

Techniques: Luciferase, Western Blot, Incubation, MTT Assay

(A) MCF-7 cells were infected with adenoviral luciferase or adenoviral elafin. DNA content was analyzed by propidium iodide staining, cell cycle distribution was calculated using the Dean-Jett-Fox model. (B-C) MCF-7 cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). (B) Lysates were subjected to western blot analysis and probed with indicated antibodies. (C) CDK-4 was immunopercipitated from 250 μg of protein lysates, the immunocomplex was then subjected to in vitro kinase assay using GST-Rb as a substrate.

Journal:

Article Title: The Neutrophil Elastase Inhibitor, Elafin, Triggers Rb-Mediated Growth Arrest and Caspase-Dependent Apoptosis in Breast Cancer

doi: 10.1158/0008-5472.CAN-10-1547

Figure Lengend Snippet: (A) MCF-7 cells were infected with adenoviral luciferase or adenoviral elafin. DNA content was analyzed by propidium iodide staining, cell cycle distribution was calculated using the Dean-Jett-Fox model. (B-C) MCF-7 cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). (B) Lysates were subjected to western blot analysis and probed with indicated antibodies. (C) CDK-4 was immunopercipitated from 250 μg of protein lysates, the immunocomplex was then subjected to in vitro kinase assay using GST-Rb as a substrate.

Techniques: Infection, Luciferase, Staining, Western Blot, In Vitro, Kinase Assay

(A) MCF-7 RBKD, shRNA control cells, MCF-7 pcDNA3.1 empty vector and pcDNA3.1-caspase 3 expressing cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). Lysates from MCF-7 pcDNA3.1 empty vector and pcDNA3.1-caspase 3 expressing cells subjected to western blot analysis 48 hours post treatment (right panels). Apoptosis was assessed by TUNEL assay (left panels). (B) Stable clones and a stable pool were generated on the MCF-7 RBKD background expressing either pcDNA3.1 empty vector or pcDNA3.1-caspase 3. Cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). Lysates were collected from each cell line and subjected to western blot analysis with the indicated antibodies. PBS treated cells were used to assess the levels of Rb and caspase 3 and MDA-MB-231 lysate was used as a control for caspase 3 expression. (C) Viability was measured by MTT assay every 24 hours for 120 hours and calculated at each time point by normalizing values from luciferase and elafin treated cells to PBS control then plotting the difference between the viability of elafin and luciferase (i.e. elafin effect – viral effect). (D-E) Stable clones and a stable pool were also generated on the MCF-7 control shRNA background expressing either pcDNA3.1 empty vector or pcDNA3.1-caspase 3. These cells were assayed in the same manner as panels B and C. (F) The viabilities measured at 120 hours in the cell lines generated from the MCF-7 RBKD and MCF-7 control shRNA as well as the parental cell lines were pooled and statistically compared. (F) Apoptosis was measured by TUNEL assay.

Journal:

Article Title: The Neutrophil Elastase Inhibitor, Elafin, Triggers Rb-Mediated Growth Arrest and Caspase-Dependent Apoptosis in Breast Cancer

doi: 10.1158/0008-5472.CAN-10-1547

Figure Lengend Snippet: (A) MCF-7 RBKD, shRNA control cells, MCF-7 pcDNA3.1 empty vector and pcDNA3.1-caspase 3 expressing cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). Lysates from MCF-7 pcDNA3.1 empty vector and pcDNA3.1-caspase 3 expressing cells subjected to western blot analysis 48 hours post treatment (right panels). Apoptosis was assessed by TUNEL assay (left panels). (B) Stable clones and a stable pool were generated on the MCF-7 RBKD background expressing either pcDNA3.1 empty vector or pcDNA3.1-caspase 3. Cells were treated with PBS (P), adenoviral luciferase (L), or adenoviral elafin (E). Lysates were collected from each cell line and subjected to western blot analysis with the indicated antibodies. PBS treated cells were used to assess the levels of Rb and caspase 3 and MDA-MB-231 lysate was used as a control for caspase 3 expression. (C) Viability was measured by MTT assay every 24 hours for 120 hours and calculated at each time point by normalizing values from luciferase and elafin treated cells to PBS control then plotting the difference between the viability of elafin and luciferase (i.e. elafin effect – viral effect). (D-E) Stable clones and a stable pool were also generated on the MCF-7 control shRNA background expressing either pcDNA3.1 empty vector or pcDNA3.1-caspase 3. These cells were assayed in the same manner as panels B and C. (F) The viabilities measured at 120 hours in the cell lines generated from the MCF-7 RBKD and MCF-7 control shRNA as well as the parental cell lines were pooled and statistically compared. (F) Apoptosis was measured by TUNEL assay.

Techniques: shRNA, Plasmid Preparation, Expressing, Luciferase, Western Blot, TUNEL Assay, Clone Assay, Generated, MTT Assay

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