NEDD1 Rabbit Polyclonal Antibody
Catalog# HA500294
NEDD1 Rabbit Polyclonal Antibody
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WB
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IHC-P
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IF-Cell
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FC
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Human
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Mouse
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unconjugated
Overview
Product Name
NEDD1 Rabbit Polyclonal Antibody
Antibody Type
Rabbit Polyclonal Antibody
Immunogen
Recombinant protein within human NEDD1 aa 1-200 / 660.
Species Reactivity
Human, Mouse
Validated Applications
WB, IHC-P, IF-Cell, FC
Target Molecular Weight
Predicted band size: 72 kDa
Positive Control
U-2 OS cell lysate, Mouse lung tissue lysate, U-2 OS, human skin tissue.
Conjugation
unconjugated
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
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), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Immunogen affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IHC-P
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1:100-1:500
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IF-Cell
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1:100
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FC
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1:1,000
Target
Function
NEDD1 (neural precursor cell expressed, developmentally down-regulated 1), also known as GCP-WD, is a homolog of the Drosophila protein known as Dgp71WD. It is a ubiquitously expressed, evolutionarily conserved protein and contains eight WD40 repeats and a coiled coil domain at the C-terminus. NEDD1 is a subunit of the γ-tubulin ring complex (γTuRC) and plays an important role in mitosis. During mitosis NEDD1 is phosphorylated and functions in forming the association of γ-tubulin with the spindle. The state of phosphorylation of NEDD1 is also important for determining its cellular localization. NEDD1 is responsible for targeting γTuRC to the centrosome and spindle and is therefore required for centrosomal and chromatin-mediated microtubule nucleation. The inhibition of NEDD1 results in the loss of γTuRC from the centrosome and a sequential loss of microtubule nucleation. Due to its critical role in mitosis, NEDD1 may be a potential target for anticancer therapies.
Background References
1. Courthéoux T. et. al. Microtubule nucleation during central spindle assembly requires NEDD1 phosphorylation on serine 405 by Aurora A. J Cell Sci. 2019 May
Subcellular Location
Centrosome.
Synonyms
FLJ35902 antibody
GCP WD antibody
NEDD-1 antibody
NEDD1 antibody
NEDD1_HUMAN antibody
Neural precursor cell expressed developmentally down-regulated protein 1 antibody
Neural precursor cell expressed, developmentally down regulated gene 1 antibody
Protein NEDD1 antibody
TUBGCP7 antibody
Images
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Western blot analysis of NEDD1 on different lysates with Rabbit anti-NEDD1 antibody (HA500294) at 1/5,000 dilution.
Lane 1: U-2 OS cell lysate
Lane 2: Mouse lung tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 72 kDa
Observed band size: 72 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 (HA500294) 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. -
Immunocytochemistry analysis of U-2 OS cells labeling NEDD1 with Rabbit anti-NEDD1 antibody (HA500294) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-NEDD1 antibody (HA500294) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-NEDD1 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA500294, 1/400) 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.
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Flow cytometric analysis of U-2 OS cells labeling NEDD1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA500294, 1/1,000) (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).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"