IRF5 Antibody

Novus Biologicals | Catalog # NB100-1092

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Predicted:

Bovine (100%), Rat (100%). Backed by our 100% Guarantee.

Applications

Validated:

Western Blot, Peptide ELISA, Flow Cytometry, Chromatin Immunoprecipitation (ChIP), Knockdown Validated

Cited:

Western Blot, Flow Cytometry, Chemotaxis

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

Peptide with sequence C-QGPWPMHPAGMQ corresponding to C-Terminus according to NP_002191.1, NP_116032.1, NP_001092097.1, NP_001092099.1.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 22879997).

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

Clonality

Polyclonal

Host

Goat

Isotype

IgG

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.

Scientific Data Images for IRF5 Antibody

Western Blot: IRF5 Antibody [NB100-1092]

Western Blot: IRF5 Antibody [NB100-1092]

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
Western Blot: IRF5 Antibody [NB100-1092]

Western Blot: IRF5 Antibody [NB100-1092]

IRF5-Antibody-Western-Blot-NB100-1092-img0003.jpg
Western Blot: IRF5 Antibody [NB100-1092]

Western Blot: IRF5 Antibody [NB100-1092]

IRF5-Antibody-Western-Blot-NB100-1092-img0002.jpg
IRF5 Antibody

Western Blot: IRF5 Antibody [NB100-1092] -

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 (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3053), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF5 Antibody

Western Blot: IRF5 Antibody [NB100-1092] -

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 (https://www.nature.com/articles/srep31002), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF5 Antibody

Western Blot: IRF5 Antibody [NB100-1092] -

Western Blot: IRF5 Antibody [NB100-1092] - Down-regulation of IRF5 protein expression by siRNAs alters sensitivity to DNA damage. A. MCF-12A cells were incubated with transfection reagent alone (mock-transfected), control Lamin A/C siRNAs or 5 nM IRF5 siRNAs once (IRF5 siRNA1) or twice (IRF5 siRNA2), as described in the Materials & methods. Western blot analysis shows > 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 (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3053), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF5 Antibody

Western Blot: IRF5 Antibody [NB100-1092] -

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 (https://www.nature.com/articles/srep31002), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF5 Antibody

Western Blot: IRF5 Antibody [NB100-1092] -

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 (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3053), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
IRF5 Antibody

Flow Cytometry: IRF5 Antibody [NB100-1092] -

Comparative analysis of IRF5 protein expression between immortalized lymphoid cell lines using flow cytometry.(A) Experimental details for the comparative analysis of alpha -IRF5 antibody specificity by flow cytometry. (B) Representative histogram plots from flow cytometry analysis of intracellular IRF5 in immortalized cell lines using different alpha -IRF5 antibodies. Data are representative of three independent experiments. Ctrl-iso, matched isotype antibodies used as non-specific controls. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27481535), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for IRF5 Antibody

Application
Recommended Usage

Peptide ELISA

Detection limit 1:128000

Western Blot

0.3 - 1 ug/mL
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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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
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  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris saline (20 mM Tris pH 7.3, 150 mM NaCl), 0.5% BSA

Preservative

0.02% Sodium Azide

Concentration

0.5 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: IRF5

Interferon regulatory factors (IRFs) are transcription factors that possess a novel helix-turn-helix DNA-binding motif and mediate virus- and interferon (IFN)-induced signaling pathways. The IRF5 protein is structurally related to IRF6. Like other IRFs, IRF5 and IRF6 contain 5 conserved tryptophan residues in the 115-amino acid N-terminal DNA-binding domain. IRF5 has a central IRF association domain (IAD) and, like IRF7 and IRF8, is induced by IFNA/IFNB stimulation. IRF5 has a type I IFN transactivation potential similar to that of IRF1. IRF3 and IRF5 form heterodimers and are both required for effective induction of IFNA gene transcription.

Long Name

Interferon Regulatory Factor 5

Alternate Names

IBD14, interferon regulatory factor 5, IRF-5, SLEB10

Entrez Gene IDs

3663 (Human)

Gene Symbol

IRF5

UniProt

Additional IRF5 Products

Product Documents for IRF5 Antibody

Certificate of Analysis

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Product Specific Notices for IRF5 Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

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Protocols

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