![Western Blot: Human IgM Antibody (P5E2) [NBP1-78603] - Analysis of 1ug of purified Human IgM, Human IgA, and Human IgG and 10ul of PageRuler Prestained Protein Ladder per well.](http://images.novusbio.com/fullsize/Human-IgM-Antibody-P5E2-Western-Blot-NBP1-78603-img0004.jpg "Western Blot: Human IgM Antibody (P5E2) [NBP1-78603] - Analysis of 1ug of purified Human IgM, Human IgA, and Human IgG and 10ul of PageRuler Prestained Protein Ladder per well.")
| Reactivity | HuSpecies Glossary |
| Applications | WB, ELISA, Flow, Func, IP |
| Clone | P5E2 |
| Clonality | Monoclonal |
| Host | Mouse |
| Conjugate | Unconjugated |
| Concentration | 1 mg/ml |
| Description | Novus Biologicals Mouse Human IgM Antibody (P5E2) (NBP1-78603) is a monoclonal antibody validated for use in WB, ELISA, Flow and IP. All Novus Biologicals antibodies are covered by our 100% guarantee. |
| Immunogen | Purified Human Serum IgM |
| Isotype | IgG1 |
| Clonality | Monoclonal |
| Host | Mouse |
| Purity | Ion exchange chromatography |
| 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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| Storage | Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
| Buffer | 0.01M potassium phosphate with 0.135M NaCl, 1mg/ml BSA, 2.5mM KCl |
| Preservative | 0.05% Sodium Azide |
| Concentration | 1 mg/ml |
| Purity | Ion exchange chromatography |
Secondary Antibodies |
Isotype Controls |
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![Flow Cytometry: Human IgM Antibody (P5E2) [NBP1-78603] - Analysis of surface IgM expression was performed on Ramos (surface and cytoplasmic IgG/lambda-expressing B cells), Raji (cytoplasmic IgG/kappa-expressing B cells), and Jurkat (T cells) cell lines. Cells were stained with a mouse anti-human IgM monoclonal antibody or mouse IgG1 isotype control (black histogram), each at a concentration of 1ug/ml. After incubation of the primary antibody on ice for 1 hour, the cells were stained with a FITC-conjugated goat anti-mouse IgG Fc secondary antibody at a dilution of 1:500 for 30 minutes on ice. The representative 10,000 cells were obtained for each sample.](http://images.novusbio.com/fullsize/Human-IgM-Antibody-P5E2-Flow-Cytometry-NBP1-78603-img0003.jpg "Flow Cytometry: Human IgM Antibody (P5E2) [NBP1-78603] - Analysis of surface IgM expression was performed on Ramos (surface and cytoplasmic IgG/lambda-expressing B cells), Raji (cytoplasmic IgG/kappa-expressing B cells), and Jurkat (T cells) cell lines. Cells were stained with a mouse anti-human IgM monoclonal antibody or mouse IgG1 isotype control (black histogram), each at a concentration of 1ug/ml. After incubation of the primary antibody on ice for 1 hour, the cells were stained with a FITC-conjugated goat anti-mouse IgG Fc secondary antibody at a dilution of 1:500 for 30 minutes on ice. The representative 10,000 cells were obtained for each sample.")
![ELISA: Human IgM Antibody (P5E2) [NBP1-78603] - Direct ELISA analysis of various immunoglobulin (Ig) isotypes was performed by coating wells of a 96-well plate with 100ul per well of Human IgM, Human IgG, or Human IgA diluted in carbonate/bicarbonate buffer, starting at a concentration of 50ng/ml and serially diluting 2-fold to a concentration of 0.05ng/ml, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 100ul per well of a mouse anti-human IgM monoclonal antibody at a concentration of 1ug/ml for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondary antibody at a dilution of 1:8000 for 1 hour at room temperature. Detection was performed using 1-Step Ultra TMB substrate for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.](http://images.novusbio.com/fullsize/Human-IgM-Antibody-P5E2-ELISA-NBP1-78603-img0001.jpg "ELISA: Human IgM Antibody (P5E2) [NBP1-78603] - Direct ELISA analysis of various immunoglobulin (Ig) isotypes was performed by coating wells of a 96-well plate with 100ul per well of Human IgM, Human IgG, or Human IgA diluted in carbonate/bicarbonate buffer, starting at a concentration of 50ng/ml and serially diluting 2-fold to a concentration of 0.05ng/ml, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 100ul per well of a mouse anti-human IgM monoclonal antibody at a concentration of 1ug/ml for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondary antibody at a dilution of 1:8000 for 1 hour at room temperature. Detection was performed using 1-Step Ultra TMB substrate for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.")
![ELISA: Human IgM Antibody (P5E2) [NBP1-78603] - Direct ELISA analysis of IgM was performed by coating wells of a 96-well plate with 100ul per well of Human IgM diluted in carbonate/bicarbonate buffer, starting at a concentration of 5ug/ml and serially diluting 2-fold to a concentration of 5ng/ml, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 100ul per well of a mouse anti-human IgM monoclonal antibody at a concentration of 1ug/ml for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondary antibody at a dilution of 1:8000 for 1 hour at room temperature. Detection was performed using 1-Step Ultra TMB substrate for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.](http://images.novusbio.com/fullsize/Human-IgM-Antibody-P5E2-ELISA-NBP1-78603-img0002.jpg "ELISA: Human IgM Antibody (P5E2) [NBP1-78603] - Direct ELISA analysis of IgM was performed by coating wells of a 96-well plate with 100ul per well of Human IgM diluted in carbonate/bicarbonate buffer, starting at a concentration of 5ug/ml and serially diluting 2-fold to a concentration of 5ng/ml, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 100ul per well of a mouse anti-human IgM monoclonal antibody at a concentration of 1ug/ml for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondary antibody at a dilution of 1:8000 for 1 hour at room temperature. Detection was performed using 1-Step Ultra TMB substrate for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.")
![SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.](https://images.novusbio.com/images/Mouse-IgG1-Isotype-Control-MG1-SDS-Page-NBP1-97005-img0002.jpg "SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.")