NMI Recombinant Rabbit Monoclonal Antibody [JE65-78]
cat.: HA720087
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: JE65-78
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 35 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human NMI aa 1-51/307.
Positive control: THP-1 cell lysate, HeLa cell lysate, K-562 cell lysate, human spleen tissue, Hela, SiHa.
Subcellular location: Nucleus, Secreted, Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC

1:1,000
1:50
1:400
1:500-1:1,000
Uniprot #: SwissProt: Q13287 Human
Alternative names: N myc (and STAT) interactor N myc and STAT interactor N myc interactor NMI NMYC and STAT interactor NMYC interactor
Images
HA720087_1.jpg Fig1: Western blot analysis of NMI on different lysates with Rabbit anti-NMI antibody (HA720087) at 1/5,000 dilution.

Lane 1: THP-1 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: K-562 cell lysate (20 µg/Lane)

Predicted band size: 35 kDa
Observed band size: 39 kDa
Exposure time: 25 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA720087) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA720087_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-NMI antibody (HA720087) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA720087) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA720087_3.jpg Fig3: Immunocytochemistry analysis of Hela cells labeling NMI with Rabbit anti-NMI antibody (HA720087) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-NMI antibody (HA720087) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) were used as the secondary antibody at 1/1,000 dilution.
HA720087_4.jpg Fig4: Immunocytochemistry analysis of SiHa cells labeling NMI with Rabbit anti-NMI antibody (HA720087) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-NMI antibody (HA720087) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) were used as the secondary antibody at 1/1,000 dilution.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.