This antibody gave a positive signal in the following tissue lysates: Mouse Brain; Mouse Hippocampus; Mouse Cerebellum; Mouse Spinal Cord; Rat Brain; Rat Spinal Cord.
This antibody gave a positive result when used in the following methanol fixed cell lines: MALME-3M
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml. Detects a band of approximately 252 kDa (predicted molecular weight: 252 kDa). Abcam recommends using milk as the blocking agent - 3%
Use a concentration of 5 µg/ml.
Proteoglycan playing a role in cell proliferation and migration which stimulates endothelial cells motility during microvascular morphogenesis. May also inhibit neurite outgrowth and growth cone collapse during axon regeneration. Cell surface receptor for collagen alpha 2(VI) which may confer cells ability to migrate on that substrate. Binds through its extracellular N-terminus growth factors, extracellular matrix proteases modulating their activity. May regulate MPP16-dependent degradation and invasion of type I collagen participating in melanoma cells invasion properties. May modulate the plasminogen system by enhancing plasminogen activation and inhibiting angiostatin. Functions also as a signal transducing protein by binding through its cytoplasmic C-terminus scaffolding and signaling proteins. May promote retraction fiber formation and cell polarization through Rho GTPase activation. May stimulate alpha-4, beta-1 integrin-mediated adhesion and spreading by recruiting and activating a signaling cascade through CDC42, ACK1 and BCAR1. May activate FAK and ERK1/ERK2 signaling cascades.
O-glycosylated; contains glycosaminoglycan chondroitin sulfate which are required for proper localization and function in stress fiber formation (By similarity). Involved in interaction with MMP16 and ITGA4. Phosphorylation by PRKCA regulates its subcellular location and function in cell motility.
Apical cell membrane. Cell projection > lamellipodium membrane. Localized at the apical plasma membrane it relocalizes to the lamellipodia of astrocytoma upon phosphorylation by PRKCA. Localizes to the retraction fibers. Localizes to the plasma membrane of oligodendrocytes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NG2 antibody (ab129051)This image is courtesy of an abreview submitted by Carl Hobbs (KCL, United Kingdom)
IHC-P image of NG2 staining of human melanoma cells in mouse tissue sections using ab129051 (1: 500). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked using 1% BSA for 10 mins at 21°C. ab129051 was dikuted 1:500 using TBS containing BSA and Azide, incubated on the sections for 2 hours at 21°C. The secondary antibody used was Goat polyclonal to Rabbit IgG conjugated to Biotin (1:250).
ICC/IF image of ab129051 stained Malme-3M cells. The cells were 100% methanol fixed (5 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab129051 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Western blot - Anti-NG2 antibody (ab129051)
All lanes : Anti-NG2 antibody (ab129051) at 1 µg/ml (Milk blocking - 3%)
Lane 1 : Brain (Mouse) Tissue Lysate Lane 2 : Mouse Hippocampus Tissue Lysate Lane 3 : Cerebellum Mouse Tissue Lysate Lane 4 : Spinal Cord (Mouse) Tissue Lysate Lane 5 : Brain (Rat) Tissue Lysate Lane 6 : Spinal Cord (Rat) Tissue Lysate
Lysates/proteins at 25 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 252 kDa Observed band size : 252 kDa
Exposure time : 20 minutesAbcam recommends using milk as the blocking agent - 3%.
This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab129051 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.