Anti-MyD88 (IN) Antibody (2125)
$445.00
| Host | Quantity | Applications | Species Reactivity | Data Sheet | |
|---|---|---|---|---|---|
| Rabbit | 100ug | ELISA,WB,IHC-P,IF,IP | Human, Mouse |  |
Overview
Product Name Anti-MyD88 (IN) Antibody (2125)
Description Anti-MYD88 (IN) Rabbit Polyclonal Antibody
Target MyD88 (IN)
Species Reactivity Human, Mouse
Applications ELISA,WB,IHC-P,IF,IP
Host Rabbit
Clonality Polyclonal
Isotype IgG
Immunogen Peptide corresponding to aa 233-248 of human MyD88. The sequence differs from mouse MyD88 by two amino acids.
Properties
Form Liquid
Concentration Lot Specific
Formulation PBS, pH 7.4.
Buffer Formulation Phosphate Buffered Saline
Buffer pH pH 7.4
Format Purified
Purification Purified by peptide immuno-affinity chromatography
Specificity Information
Specificity This antibody recognizes human and mouse MyD88 (35 kD).
Target Name Myeloid differentiation primary response protein MyD88
Target ID MyD88 (IN)
Uniprot ID Q99836
Gene Name MYD88
Gene ID 4615
Accession Number NP_002459
Sequence Location Cytoplasm, Nucleus
Biological Function Adapter protein involved in the Toll-like receptor and IL-1 receptor signaling pathway in the innate immune response (PubMed:15361868, PubMed:18292575, PubMed:33718825). Acts via IRAK1, IRAK2, IRF7 and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response (PubMed:15361868, PubMed:24316379, PubMed:19506249). Increases IL-8 transcription (PubMed:9013863). Involved in IL-18-mediated signaling pathway. Activates IRF1 resulting in its rapid migration into the nucleus to mediate an efficient induction of IFN-beta, NOS2/INOS, and IL12A genes. Upon TLR8 activation by GU-rich single-stranded RNA (GU-rich RNA) derived from viruses such as SARS-CoV-2, SARS-CoV and HIV-1, induces IL1B release through NLRP3 inflammasome activation (PubMed:33718825). MyD88-mediated signaling in intestinal epithelial cells is crucial for maintenance of gut homeostasis and controls the expression of the antimicrobial lectin REG3G in the small intestine (By similarity). {UniProtKB:P22366, PubMed:15361868, PubMed:18292575, PubMed:19506249, PubMed:20855887, PubMed:24316379, PubMed:33718825, PubMed:9013863}.
Research Areas Neuroscience
Background Cellular responses induced by the pro- inflammatory cytokine IL-1 require IL-1 receptor complex (IL-1R1 and IL- 1RacP). Recently, MyD88 was identified as an adapter molecule in the IL-1 signaling pathway. MyD88 associates with and recruits IRAK to the IL-1 receptor. Dominant negative mutants of MyD88 attenuate IL-1R- mediated NF-kappaB activation. MyD88 also functions as a regulator molecule for IL- 18 receptor and human Toll receptor, members of the Toll/IL-1R family of receptors. Targeted disruption of the MyD88 gene results in loss of cellular responses to IL-1 and IL-18, and MyD88-deficient mice lack responses to LPS which require Toll-like receptors 2 and 4 (TLR2 and TLR4) as the signaling receptors. MyD88 is a general adapter protein for the Toll/IL-1R family of receptors and plays an important role in the inflammatory responses induced by cytokines IL-1, IL-18, and LPS. MyD88 is expressed in a variety of tissues.
Application Images
Description Western Blot Validation of MyD88 in HeLa (A) and Jurket (B) Cells
Loading: 15 ug of lysates per lane. Antibodies: 2125 (1 ug/mL) 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: 15 ug of lysates per lane. Antibodies: 2125 (1 ug/mL) 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 15 ug of lysates per lane. Antibodies: MyD88 2125 (2 ug/mL), MyD88 2127 (2 ug/mL), beta-actin (1 ug/mL), and GAPDH (0.02 ug/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: 15 ug of lysates per lane. Antibodies: MyD88 2125 (2 ug/mL), MyD88 2127 (2 ug/mL), beta-actin (1 ug/mL), and GAPDH (0.02 ug/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Independent Antibody Validation (IAV) via Protein Expression Profile in Human Tissues
Loading: 15 ug of lysates per lane. Antibodies: MyD88 2125 (2 ug/mL), MyD88 2127 (2 ug/mL), beta-actin (1 ug/mL), and GAPDH (0.02 ug/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: 15 ug of lysates per lane. Antibodies: MyD88 2125 (2 ug/mL), MyD88 2127 (2 ug/mL), beta-actin (1 ug/mL), and GAPDH (0.02 ug/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Animal Species Reactivity
Loading: Lysates/proteins at 15 ug per lane. Antibodies: 2125 (2 ug/mL) or 2127 (2 ug/mL). 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Loading: Lysates/proteins at 15 ug per lane. Antibodies: 2125 (2 ug/mL) or 2127 (2 ug/mL). 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Validation with MyD88 siRNA Knockdown in HeLa Cells
HeLa cells were transfected with control siRNAs (lane 1) or MyD88 siRNAs (lane 2) Loading: 10 ug of HeLa whole cell lysates per lane. Antibodies: 2125 (2 ug/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
HeLa cells were transfected with control siRNAs (lane 1) or MyD88 siRNAs (lane 2) Loading: 10 ug of HeLa whole cell lysates per lane. Antibodies: 2125 (2 ug/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Description Immunofluorescence Validation of MyD88 n Human Testis
Immunofluorescent analysis of 4% paraformaldehyde-fixed human testis tissue labeling MyD88 with 2125 at 20 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing nucleus staining on human testis cells.
Immunofluorescent analysis of 4% paraformaldehyde-fixed human testis tissue labeling MyD88 with 2125 at 20 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing nucleus staining on human testis cells.
Description Immunohistochemistry Validation of MyD88 in Human Heart
Immunohistochemical analysis of paraffin-embedded human heart tissue using anti-MyD88 antibody (2125) at 2 ug/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
Immunohistochemical analysis of paraffin-embedded human heart tissue using anti-MyD88 antibody (2125) at 2 ug/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
Description Immunofluorescence Validation of MyD88 in K562 Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling MyD88 with 2125 at 10 ug/mL, followed by Goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling MyD88 with 2125 at 10 ug/mL, followed by Goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
Description KO Validation in Mouse Macrophages (Miller et al., 2006)
Bone marrow-derived macrophages from wild type (WT) mice and MyD88 knockout mice were assessed for MyD88 protein expression by anti-MyD88 antibodies. MyD88 expression was detected in WT mice, but not in MyD88 knockout mice.
Bone marrow-derived macrophages from wild type (WT) mice and MyD88 knockout mice were assessed for MyD88 protein expression by anti-MyD88 antibodies. MyD88 expression was detected in WT mice, but not in MyD88 knockout mice.
Description KO Validation in MyD88-deficient MEF cell line (Burns et al., 2424)
MyD88−/− deficient MEF cell line was reconstituted by retroviral infection with an empty vector, MyD88, or MyD88s expression vectors. The levels of MyD88 (isoform 1) or MyD88s (isoform 3) were confirmed with anti-MyD88 antibodies and MyD88 expression was not detected in the MyD88-deficient cells.
MyD88−/− deficient MEF cell line was reconstituted by retroviral infection with an empty vector, MyD88, or MyD88s expression vectors. The levels of MyD88 (isoform 1) or MyD88s (isoform 3) were confirmed with anti-MyD88 antibodies and MyD88 expression was not detected in the MyD88-deficient cells.
Description KO Validation in Mouse IECs (Vlantis et al., 2014)
Western blot with anti-MyD88 antibodies on intestinal epithelial cells (IECs) showing efficient deletion of MyD88 and concomitant expression of GFP in MyD88IEC-KO mice, but not in MyD88 knockout mice.
Western blot with anti-MyD88 antibodies on intestinal epithelial cells (IECs) showing efficient deletion of MyD88 and concomitant expression of GFP in MyD88IEC-KO mice, but not in MyD88 knockout mice.
Description KD Validation in Chondrocytes (Ahmad et al., 2009)
Chondrocytes were transfected with either MyD88 siRNA or control siRNA and analyzed for MyD88 expression by immunoblotting with anti-Myd88 antibodies that confirmed inhibition of the target proteins.
Chondrocytes were transfected with either MyD88 siRNA or control siRNA and analyzed for MyD88 expression by immunoblotting with anti-Myd88 antibodies that confirmed inhibition of the target proteins.
Description Immunoprecipitation Validation in HEK293 cells (Kawai et al., 2004)
HEK293 cells were transiently transfected with FLAG-IRF7. Cell lysates were immunoprecipitated with control rabbit anti-mouse immunoglobulin serum (IgG) or anti-MyD88 (Ab1 and Ab2), followed by immunoblotting with anti-FLAG.
HEK293 cells were transiently transfected with FLAG-IRF7. Cell lysates were immunoprecipitated with control rabbit anti-mouse immunoglobulin serum (IgG) or anti-MyD88 (Ab1 and Ab2), followed by immunoblotting with anti-FLAG.
Handling
Storage This antibody is stable for at least one (1) year at -20°C. Avoid multiple freeze- thaw cycles.
Dilution Instructions Dilute in PBS or medium which is identical to that used in the assay system.
Application Instructions Immunoblotting: use at 1:500-1:1,000 dilution.
Positive control: Whole cell lysate from Jurkat cells.
Positive control: Whole cell lysate from Jurkat cells.
References & Data Sheet
Data Sheet 
Download PDF Data Sheet
Download PDF Data Sheet