![Western Blot: IRF5 Antibody [NB100-1092] - Western blot analysis of Human Spleen (A) and A549 cell lysate (B) + peptide (C), 35 ug protein in RIPA buffer. Antibody at 1 ug/mL. Detected by chemiluminescence](http://images.novusbio.com/fullsize/IRF5-Antibody-Western-Blot-NB100-1092-img0004.jpg "Western Blot: IRF5 Antibody [NB100-1092] - Western blot analysis of Human Spleen (A) and A549 cell lysate (B) + peptide (C), 35 ug protein in RIPA buffer. Antibody at 1 ug/mL. Detected by chemiluminescence")
| Reactivity | Hu, Mu, Rt, BvSpecies Glossary |
| Applications | WB, ELISA, Flow, ChIP, KD |
| Clonality | Polyclonal |
| Host | Goat |
| Conjugate | Unconjugated |
| Concentration | 0.5 mg/ml |
| Immunogen | Peptide with sequence C-QGPWPMHPAGMQ corresponding to C-Terminus according to NP_002191.1, NP_116032.1, NP_001092097.1, NP_001092099.1. |
| Specificity | This antibody is expected to recognize all four reported isoforms (NP_002191.1; NP_116032.1; NP_001092097.1; NP_001092099.1). Reported variants NP_001092100.1, NP_001092101.1, NP_001092098.1, NP_001092097.1 represent identical protein: Reported variants NP_116032.1, NP_001092100.1 represent identical protein |
| Predicted Species | Rat (100%), Bovine (100%). Backed by our 100% Guarantee. |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Goat |
| Gene | IRF5 |
| Purity | Immunogen affinity purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Dilutions |
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| Application Notes | Western blot: Approx 55+60 kDa bands observed in Human spleen lysates and approx 65 kDa in lysates of cell line A549 (calculated MW of 57.8 kDa according to NP_001092099.1 and 56.0 kDa according to NP_116032.1; NP_001092097.2). Use in ChIP reported in scientific literature (PMID: 20237317). Use in Flow cytometry reported in scientific literature (PMID: 22879997). |
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| Theoretical MW | 56 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
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| Control |
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| Publications |
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| Storage | Store at -20C. Avoid freeze-thaw cycles. |
| Buffer | Tris saline (20 mM Tris pH 7.3, 150 mM NaCl), 0.5% BSA |
| Preservative | 0.02% Sodium Azide |
| Concentration | 0.5 mg/ml |
| Purity | Immunogen affinity purified |
Secondary Antibodies |
Isotype Controls |
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![Western Blot: IRF5 Antibody [NB100-1092] - Endogenous IRF5 knockdown analysis. Ramos B cells were nucleofected with scrambled (Scr) or IRF5 (KD) siRNAs and lysates from same nucleofection run on multiple independent blots for comparative analysis of seven different commercially available alpha-IRF5 antibodies. Arrows indicate detection of an appropriately sized band(s) corresponding to IRF5. Data are representative of three independent experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep31002), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/IRF5-Antibody-Western-Blot-NB100-1092-img0003.jpg "Western Blot: IRF5 Antibody [NB100-1092] - Endogenous IRF5 knockdown analysis. Ramos B cells were nucleofected with scrambled (Scr) or IRF5 (KD) siRNAs and lysates from same nucleofection run on multiple independent blots for comparative analysis of seven different commercially available alpha-IRF5 antibodies. Arrows indicate detection of an appropriately sized band(s) corresponding to IRF5. Data are representative of three independent experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep31002), licensed under a CC-BY license.")
![Western Blot: IRF5 Antibody [NB100-1092] - Comparative analysis of IRF5 protein expression between immortalized lymphoid cell lines by immunoblot. Endogenous IRF5 protein expression in lysates (T, THP-1; R, Ramos B cells; J, Jurkat T cells) was detecting by running multiple independent blots for the analysis of antibody specificity; seven different commercially available alpha-IRF5 antibodies were evaluated. IRF5 expression was detected with the previously validated Cell Signaling antibody #3257 that is no longer available. beta-actin levels and apparent molecular weight standards are shown as loading controls and for size comparison, respectively. Arrows indicate detection of an appropriately sized band(s) corresponding to IRF5. Data are representative of three independent experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep31002), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/IRF5-Antibody-Western-Blot-NB100-1092-img0002.jpg "Western Blot: IRF5 Antibody [NB100-1092] - Comparative analysis of IRF5 protein expression between immortalized lymphoid cell lines by immunoblot. Endogenous IRF5 protein expression in lysates (T, THP-1; R, Ramos B cells; J, Jurkat T cells) was detecting by running multiple independent blots for the analysis of antibody specificity; seven different commercially available alpha-IRF5 antibodies were evaluated. IRF5 expression was detected with the previously validated Cell Signaling antibody #3257 that is no longer available. beta-actin levels and apparent molecular weight standards are shown as loading controls and for size comparison, respectively. Arrows indicate detection of an appropriately sized band(s) corresponding to IRF5. Data are representative of three independent experiments. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep31002), licensed under a CC-BY license.")
![Western Blot: IRF5 Antibody [NB100-1092] - Overexpression of IRF5 in MCF-7 & MDA-MB-231 cells sensitizes them to DNA damage-induced growth inhibition. A. Endogenous IRF expression was analyzed by Western blot in transformed mammary epithelial cell lines. Levels of beta -actin are shown as loading controls. B. Western blot analysis of stable cell lines generated to overexpress retroviral pBIRF5. C. Cell survival was measured in MCF-7 & MDA-MB-231 pBabe cell lines by colony formation assay before & after treatment. Cells were treated with 0.1 or 1 μM Doxorubicin (Dox) or 2, 5 & 10 Gy gamma -IR. The number of colonies is plotted on the y-axis as percent of control; 100% represents the number of colonies in empty pBabe control lines. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between colonies in pBabe versus pBIRF5 cell lines after each treatment; * denotes P 70% reduction of endogenous IRF5 proteins after normalization to beta -actin levels. B. Cells were exposed to 5 Gy IR or the same dose plus IFN-gamma (IR/ gamma ) for 24 h. Percent of Annexin V-FITC stained positive cells is shown in the upper & lower right-hand quadrants. Representative histogram plots from three independent experiments performed in duplicate are shown. C. Same as in B, except cells were exposed to 1 μM Dox or Dox & IFN-gamma for five hours. Percent of Annexin V-FITC stained positive cells compared to control is plotted on y-axis. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between cells transfected with Lamin A/C siRNAs (12Asicon) & IRF5 siRNAs (12AsiIRF5) after each treatment; ** denotes P < 0.001. D. Cells were treated with the indicated doses of Dox or IR after siRNA transfection. Number of colonies is plotted on y-axis as percent of control. A total of 100% represents the number of colonies in control untreated 12Asicon cells. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between colonies in 12Asicon versus 12AsiIRF5 cells after each treatment; * denotes P < 0.05. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3053), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1092_goat-polyclonal-irf5-antibody-imgenex-img-3525-31020241612691.jpg "Western Blot: IRF5 Antibody [NB100-1092] - Overexpression of IRF5 in MCF-7 & MDA-MB-231 cells sensitizes them to DNA damage-induced growth inhibition. A. Endogenous IRF expression was analyzed by Western blot in transformed mammary epithelial cell lines. Levels of beta -actin are shown as loading controls. B. Western blot analysis of stable cell lines generated to overexpress retroviral pBIRF5. C. Cell survival was measured in MCF-7 & MDA-MB-231 pBabe cell lines by colony formation assay before & after treatment. Cells were treated with 0.1 or 1 μM Doxorubicin (Dox) or 2, 5 & 10 Gy gamma -IR. The number of colonies is plotted on the y-axis as percent of control; 100% represents the number of colonies in empty pBabe control lines. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between colonies in pBabe versus pBIRF5 cell lines after each treatment; * denotes P 70% reduction of endogenous IRF5 proteins after normalization to beta -actin levels. B. Cells were exposed to 5 Gy IR or the same dose plus IFN-gamma (IR/ gamma ) for 24 h. Percent of Annexin V-FITC stained positive cells is shown in the upper & lower right-hand quadrants. Representative histogram plots from three independent experiments performed in duplicate are shown. C. Same as in B, except cells were exposed to 1 μM Dox or Dox & IFN-gamma for five hours. Percent of Annexin V-FITC stained positive cells compared to control is plotted on y-axis. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between cells transfected with Lamin A/C siRNAs (12Asicon) & IRF5 siRNAs (12AsiIRF5) after each treatment; ** denotes P < 0.001. D. Cells were treated with the indicated doses of Dox or IR after siRNA transfection. Number of colonies is plotted on y-axis as percent of control. A total of 100% represents the number of colonies in control untreated 12Asicon cells. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between colonies in 12Asicon versus 12AsiIRF5 cells after each treatment; * denotes P < 0.05. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3053), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: IRF5 Antibody [NB100-1092] - Comparative analysis of IRF5 protein expression between immortalized lymphoid cell lines by immunoblot.(A) qRT-PCR analysis of IRF5 transcript expression in immortalized lymphoid cell lines. Data is presented as relative expression after normalization to beta -actin using the ∆∆Ct method. (B) Immunoblot analysis of endogenous IRF5 protein expression in immortalized cell lines. IRF5 expression was detected with the previously validated Cell Signaling antibody #3257 that is no longer available. beta -actin levels & apparent molecular weight standards are shown as loading controls & for size comparison, respectively. (C) Experimental details for the comparative analysis of alpha -IRF5 antibody specificity by immunoblot analysis. (D) Same as in (B)except lysates (T, THP-1; R, Ramos B cells; J, Jurkat T cells) were run on multiple independent blots for the analysis of antibody specificity; seven different commercially available alpha -IRF5 antibodies were evaluated. (E) Same as in (D) except antibodies were evaluated by IRF5 knockdown analysis. Ramos B cells were nucleofected with scrambled (Scr) or IRF5 (KD) siRNAs & lysates from same nucleofection run on multiple independent blots for comparative analysis. Arrows indicate detection of an appropriately sized band(s) corresponding to IRF5. Data (except in (C)) are representative of three independent experiments. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep31002), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1092_goat-polyclonal-irf5-antibody-imgenex-img-3525-310202415525278.jpg "Western Blot: IRF5 Antibody [NB100-1092] - Comparative analysis of IRF5 protein expression between immortalized lymphoid cell lines by immunoblot.(A) qRT-PCR analysis of IRF5 transcript expression in immortalized lymphoid cell lines. Data is presented as relative expression after normalization to beta -actin using the ∆∆Ct method. (B) Immunoblot analysis of endogenous IRF5 protein expression in immortalized cell lines. IRF5 expression was detected with the previously validated Cell Signaling antibody #3257 that is no longer available. beta -actin levels & apparent molecular weight standards are shown as loading controls & for size comparison, respectively. (C) Experimental details for the comparative analysis of alpha -IRF5 antibody specificity by immunoblot analysis. (D) Same as in (B)except lysates (T, THP-1; R, Ramos B cells; J, Jurkat T cells) were run on multiple independent blots for the analysis of antibody specificity; seven different commercially available alpha -IRF5 antibodies were evaluated. (E) Same as in (D) except antibodies were evaluated by IRF5 knockdown analysis. Ramos B cells were nucleofected with scrambled (Scr) or IRF5 (KD) siRNAs & lysates from same nucleofection run on multiple independent blots for comparative analysis. Arrows indicate detection of an appropriately sized band(s) corresponding to IRF5. Data (except in (C)) are representative of three independent experiments. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep31002), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: IRF5 Antibody [NB100-1092] - Overexpression of IRF5 in MCF-7 & MDA-MB-231 cells sensitizes them to DNA damage-induced growth inhibition. A. Endogenous IRF expression was analyzed by Western blot in transformed mammary epithelial cell lines. Levels of beta -actin are shown as loading controls. B. Western blot analysis of stable cell lines generated to overexpress retroviral pBIRF5. C. Cell survival was measured in MCF-7 & MDA-MB-231 pBabe cell lines by colony formation assay before & after treatment. Cells were treated with 0.1 or 1 μM Doxorubicin (Dox) or 2, 5 & 10 Gy gamma -IR. The number of colonies is plotted on the y-axis as percent of control; 100% represents the number of colonies in empty pBabe control lines. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between colonies in pBabe versus pBIRF5 cell lines after each treatment; * denotes P < 0.05, ** P < 0.001. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3053), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-1092_goat-polyclonal-irf5-antibody-imgenex-img-3525-31020241553200.jpg "Western Blot: IRF5 Antibody [NB100-1092] - Overexpression of IRF5 in MCF-7 & MDA-MB-231 cells sensitizes them to DNA damage-induced growth inhibition. A. Endogenous IRF expression was analyzed by Western blot in transformed mammary epithelial cell lines. Levels of beta -actin are shown as loading controls. B. Western blot analysis of stable cell lines generated to overexpress retroviral pBIRF5. C. Cell survival was measured in MCF-7 & MDA-MB-231 pBabe cell lines by colony formation assay before & after treatment. Cells were treated with 0.1 or 1 μM Doxorubicin (Dox) or 2, 5 & 10 Gy gamma -IR. The number of colonies is plotted on the y-axis as percent of control; 100% represents the number of colonies in empty pBabe control lines. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between colonies in pBabe versus pBIRF5 cell lines after each treatment; * denotes P < 0.05, ** P < 0.001. Image collected & cropped by CiteAb from the following publication (//breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3053), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
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