MYD88 Antibody

• Catalog Number: orb1239647
• Category: Antibodies
• Description: MYD88 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IF, IHC-P, IP, WB
• Predicted Reactivity: Bovine, Gallus, Porcine, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Anti-MYD88 antibody (orb1239647) was raised against a peptide corresponding to 16 amino acids near the center of human MYD88 isoform 1. The immunogen is located within amino acids 220 - 270 of MYD88.
• Concentration: 1 mg/mL
• Dilution range: WB: 1 - 2 μg/mL; IHC-P: 2 μg/mL; IF: 10-20 μg/mL. Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry, Immunoprecipitation and Immunofluorescence in human samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 35kD Observed: 35kD
• Target: MYD88
• UniProt ID: U70451
• NCBI: AAB49967.1
• Storage: MYD88 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: MYD88 Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: MYD88 Antibody: Myeloid differentiation primary re
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western Blot Validation of MyD88 in HeLa (A) and Jurket (B) Cells. Loading: 15 µg of lysates per lane. Antibodies: orb1239647 (1 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 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.

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.

Validation with MyD88 siRNA Knockdown in HeLa Cells. HeLa cells were transfected with control siRNAs (lane 1) or MyD88 siRNAs (lane 2) Loading: 10 µg of HeLa whole cell lysates per lane. Antibodies: orb1239647 (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 n Human Testis. Immunofluorescent analysis of 4% paraformaldehyde-fixed human testis tissue labeling MyD88 with orb1239647 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). Image showing nucleus staining on human testis cells.

Immunohistochemistry Validation of MyD88 in Human Heart. Immunohistochemical analysis of paraffin-embedded human heart tissue using anti-MyD88 antibody (orb1239647) 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.

Immunofluorescence Validation of MyD88 in K562 Cells. Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling MyD88 with orb1239647 at 10 µg/mL, followed by Goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

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.

KO Validation in MyD88-deficient MEF cell line (Burns et al., 2003). 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.

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.

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.

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.