MYD88 Antibody

• Catalog Number: orb1239635
• Category: Antibodies
• Description: MYD88 Antibody
• Target: MYD88
• Clonality: Polyclonal
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Bovine, Gallus, Porcine, Sheep
• Form/Appearance: Liquid
• Concentration: 1 mg/mL
• Buffer/Preservatives: MYD88 Antibody is supplied in PBS containing 0.02% sodium azide.
• Purification: MYD88 Antibody is affinity chromatography purified via peptide column.
• Immunogen: Anti-MYD88 antibody (orb1239635) was raised against a peptide corresponding to 17 amino acids near carboxy terminus of human MYD88 isoform 1. The immunogen is located within the last 50 amino acids of MYD88.
• UniProt ID: U70451
• MW: Predicted: 35kD Observed: 35kD
• Tested applications: ELISA, IF, IP, WB
• Antibody Type: Primary Antibody
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: MYD88 Antibody: MYD88D
• Research Area: Immunology & Inflammation, Signal Transduction
• Note: For research use only
• NCBI: AAB49967.1

Animal Species Reactivity. Loading: Lysates/proteins at 15 µg per lane. Antibodies: orb1239647 (2 µg/mL) or orb1239635 (2 µg/mL). 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunofluorescence Validation of MyD88 In HeLa Cells. Immunofluorescent analysis of methanol-fixed HeLa cells labeling MyD88 with orb1239635 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and Hoechst staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.

Immunofluorescence Validation of MyD88 in Jurkat Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed Jurkat cells labeling MyD88 with orb1239635 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

Immunohistochemistry Validation of MyD88 in Human Colon. Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-MYD88 antibody (orb1239635) at 1 µg/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 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry Validation of MyD88 in Mouse Heart. Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-MYD88 antibody (orb1239635) at 2 µg/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 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunohistochemistry Validation of MyD88 in Rat Liver. Immunohistochemical analysis of paraffin-embedded rat liver tissue using anti-MYD88 antibody (orb1239635) at 1 µg/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 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunoprecipitation Validation in HEK293 cells. 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.

Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: MyD88 orb1239647 (2 µg/mL), MyD88 orb1239635 (2 µg/mL), beta-actin (1 µg/mL), and GAPDH (0.02 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Independent Antibody Validation (IAV) via Protein Expression Profile in Human Tissues. Loading: 15 µg of lysates per lane. Antibodies: MyD88 orb1239647 (2 µg/mL), MyD88 orb1239635 (2 µg/mL), beta-actin (1 µg/mL), and GAPDH (0.02 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

KD Validation in Raw 264.7 cells. The Transfection of RAW 264.7 cells with MyD88-specific siRNA resulted in attenuation of MyD88 protein by Western blot analysis with anti-Myd88 antibodies (orb1239635).

KO Validation in HeLa Cells. Loading: 10 µg of HeLa WT cell lysate or MyD88 KO cell lysate. Antibodies: MyD88 orb1239635 (2 µg/mL) and beta-actin orb1240312 (1 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

KO Validation of MyD88 in Mouse T cells. Splenocytes were isolated from a Myd88ΔT mouse in which MyD88 was specifically disrupted in T cells. T and B cells were FACS purified, and MyD88 expression was examined by Western blot with anti-MyD88 antibodies (orb1239635). MyD88 expression was detected in Splenocytes and B cells, but not in T cells.

Western Blot Validation of MyD88 in human cell lines. Loading: 15 µg of lysates per lane. Antibodies: orb1239635 (2 µg/mL) 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Predicted band size: 35 kDa.