NDUFB9 Recombinant Rabbit Monoclonal Antibody [JE63-09]
cat.: HA721015
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: JE63-09
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
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: 22 kDa
Isotype: IgG
Immunogen: Recombinant protein within human NDUFB9 aa 30-179/179.
Positive control: HepG2 cell lysate, Jurkat cell lysate, 293T cell lysate, mouse heart tissue lysate, mouse brain tissue lysate, mouse kidney tissue lysate, rat heart tissue lysate, rat kidney tissue, human liver carcinoma tissue, human stomach tissue, mouse heart tissue, PC-3M, HeLa.
Subcellular location: Mitochondrion inner membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500
1:100
1:100-1:400
1:500-1:1,000
Uniprot #: SwissProt: Q9Y6M9 Human | Q9CQJ8 Mouse
Entrez Gene: 299954 Rat
Alternative names: B22 CI B22 CI-B22 complex I B22 subunit Complex I-B22 DKFZp566O173 FLJ22885 I B22 LYR motif containing protein 3 LYR motif-containing protein 3 LYRM3 NADH dehydrogenase (ubiquinone) 1 beta subcomplex, 9, 22kDa NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 9 NADH ubiquinone oxidoreductase B22 subunit NADH-ubiquinone oxidoreductase B22 subunit NDUB9_HUMAN Ndufb9 UQOR22
Images
HA721015_1.jpg Fig1: Western blot analysis of NDUFB9 on different lysates with Rabbit anti-NDUFB9 antibody (HA721015) at 1/500 dilution.

Lane 1: HepG2 cell lysate
Lane 2: Jurkat cell lysate
Lane 3: 293T cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 22 kDa
Observed band size: 22 kDa

Exposure time: 2 minutes;

12% 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 (HA721015) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
HA721015_2.jpg Fig2: Western blot analysis of NDUFB9 on different lysates with Rabbit anti-NDUFB9 antibody (HA721015) at 1/500 dilution.

Lane 1: Mouse heart tissue lysate
Lane 2: Mouse brain tissue lysate
Lane 3: Mouse kidney tissue lysate
Lane 4: Rat heart tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 22 kDa
Observed band size: 22 kDa

Exposure time: 1 minute;

12% 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 (HA721015) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
HA721015_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-NDUFB9 antibody (HA721015) at 1/400 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 (HA721015) at 1/400 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.
HA721015_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue with Rabbit anti-NDUFB9 antibody (HA721015) at 1/400 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 (HA721015) at 1/400 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.
HA721015_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-NDUFB9 antibody (HA721015) at 1/100 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 (HA721015) at 1/100 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.
HA721015_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-NDUFB9 antibody (HA721015) at 1/100 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 (HA721015) at 1/100 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.
HA721015_7.jpg Fig7: Flow cytometric analysis of PC-3M cells labeling NDUFB9.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721015, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA721015_8.jpg Fig8: Immunocytochemistry analysis of HeLa cells labeling NDUFB9 with Rabbit anti-NDUFB9 antibody (HA721015) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NDUFB9 antibody (HA721015) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.