MA5-49239

antibody from Invitrogen Antibodies

Targeting: NMI

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-49239

Provider

Invitrogen Antibodies

Product name

NMI Monoclonal Antibody (2F3)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

Adding 0.2 mL of distilled water will yield a concentration of 500 µg/mL. Human NMI shares 64% amino acid (aa) sequence identity with mouse NMI. Positive Control - WB: human placenta tissue, K562 whole cell, Hela whole cell, A431 whole cell, PC-3 whole cell, Caco-2 whole cell, HEK293 whole cell, mouse intestine tissue, rat liver tissue. IHC: human lung cancer tissue, human colon cancer tissue, human placenta tissue. ICC/IF: SiHa cell. Flow: A431 cell.|Store at -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freeze-thaw cycles.

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

2F3

Vial size

100 µg

Concentration

500 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of NMI using NMI antibody (Product # MA5-49239). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 µg of sample under reducing conditions. Lane 1: human placenta tissue lysates; Lane 2: K562 whole cell lysates; Lane 3: Hela whole cell lysates; Lane 4: A431 whole cell lysates; Lane 5: PC-3 whole cell lysates; Lane 6: Caco-2 whole cell lysates; Lane 7: HEK293 whole cell lysates; Lane 8: mouse intestine tissue lysates; Lane 9: rat liver tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-NMI antigen affinity purified monoclonal antibody (Product # MA5-49239) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:5,000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for NMI at approximately 40KD. The expected band size for NMI is at 35KD.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of NMI in SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. Samples were then incubated in NMI Monoclonal antibody (Product # MA5-49239) using a dilution of 5 μg/mL. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NMI in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with NMI Monoclonal antibody (Product # MA5-49239) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NMI in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with NMI Monoclonal antibody (Product # MA5-49239) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NMI in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with NMI Monoclonal antibody (Product # MA5-49239) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NMI in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with NMI Monoclonal antibody (Product # MA5-49239) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of NMI in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. Samples were incubated with NMI Monoclonal antibody (Product # MA5-49239) using a dilution of 1 μg/mL overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of NMI in A431 cells (Blue line). The cells were blocked with 10% normal goat serum, and then incubated with NMI monoclonal antibody (Product # MA5-49239) (1 μg/1x10^6 cells) for 30 min at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x10^6 cells) used under the same conditions. Unlabeled sample (Red line) was also used as a control. DyLight®488 conjugated goat anti-mouse IgG (5-10 μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C.