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Product Name
Anti-NDUFB8 antibody
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Description
Rabbit monoclonal antibody to NDUFB8
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Tested applications
WB, IP, IHC-P, FC, ICC
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Species reactivity
Human, Mouse, Rat
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Alternative names
ASHI antibody; CI-ASHI antibody; MC1DN32 antibody
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Isotype
Rabbit IgG
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Preparation
This antigen of this antibody was recombinant protein within n-terminal human ndufb8 .
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Clonality
Monoclonal
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Formulation
Liquid, 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
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Storage instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
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Applications
WB: 1:500-1:2,000
IP: 1:10-1:50
IHC-P: 1:50-1:200
FC: 1:50-1:100
ICC: 1:50
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Validations
Fig1:; Western blot analysis of NDUFB8 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody ( 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.; Positive control:; Lane 1: 293 cell lysate; Lane 2: A549 cell lysate; Lane 3: Mouse heart tissue lysate; Lane 4: Rat spleen tissue lysate
Fig2:; Immunohistochemical analysis of paraffin-embedded rat heart tissue using anti-NDUFB8 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes 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.
Fig3:; Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-NDUFB8 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes 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.
Fig4:; Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-NDUFB8 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes 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.
Fig5:; Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-NDUFB8 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH; 2; O and PBS, and then probed with the primary antibody ( 1/50) for 30 minutes 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.
Fig6:; Flow cytometric analysis of NDUFB8 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody ( 1/50) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; yellow).
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References
- Stroud D A et al. Accessory subunits are integral for assembly and function of human mitochondrial complex I. Nature 538:123-126 (2016).
- Murray J et al. The subunit composition of the human NADH dehydrogenase obtained by rapid one-step immunopurification. J Biol Chem 278:13619-13622 (2003).
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