Name: Anti-NG2 antibody [EPR23976-145]
SKU: ab275024
Rating: 5 (1 reviews)

Anti-NG2 antibody [EPR23976-145] ab275024 is a rabbit monoclonal antibody that is used in NG2 western blotting, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests

Images

Flow Cytometry - Anti-NG2 antibody [EPR23976-145] (AB275024), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	Flow Cyt	WB	IHC-P
Human	Tested	Expected	Tested	Tested	Not recommended
Mouse	Tested	Tested	Tested	Tested	Not recommended
Rat	Tested	Expected	Expected	Tested	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes -
Species Rat	Dilution info 1/50	Notes -
Species Human	Dilution info 1/50	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Target data

See full target information CSPG4

Function

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.

Alternative names

MCSP, CSPG4, Chondroitin sulfate proteoglycan 4, Chondroitin sulfate proteoglycan NG2, Melanoma chondroitin sulfate proteoglycan, Melanoma-associated chondroitin sulfate proteoglycan

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR23976-145
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

Flow Cytometry - Anti-NG2 antibody [EPR23976-145] (ab275024)
Flow cytometric analysis of Mouse primary neural glia cell cells labelling NG2 with ab275024 at 1/500 dilution (0.1ug) (Right) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Left).
Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
Immunocytochemistry/ Immunofluorescence - Anti-NG2 antibody [EPR23976-145] (ab275024)
ab275024 staining NG2 in A375 cells, with negative expression in MCF7 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab275024 at 5 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
This product also work with 4% formaldehyde (10 min) fixation under the same testing conditions.
Immunocytochemistry/ Immunofluorescence - Anti-NG2 antibody [EPR23976-145] (ab275024)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neural/glia cell cells labelling NG2 with ab275024 at 1/100 (4.67 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in mouse primary glia cells. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. is observed. Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain MAP2 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).
ve control 1: ab275024 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Immunoprecipitation - Anti-NG2 antibody [EPR23976-145] (ab275024)
NG2 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab275024 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275024 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/1000 dilution.
Lane 1: Mouse brain tissue lysate 10 ug
Lane 2: ab275024 IP in Mouse brain tissue lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab275024 in mouse brain tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds.
Sample loaded onto lane 1 was non-boiled as boiling may cause protein aggregates.
All lanes: Immunoprecipitation - Anti-NG2 antibody [EPR23976-145] (ab275024)
Predicted band size: 251 kDa
Observed band size: 280 kDa, 330 kDa
Western blot - Anti-NG2 antibody [EPR23976-145] (ab275024)
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-3: 59 seconds; Lane 4: 81 seconds.
The band of 330KDa represents the intact NG2 proteoglycan modified by chondroitin sulfate, the band of 280KDa represents NG2 core protein.
The molecular weight observed is consistent with what has been described in the literature (PMID: 20455858, 16625365, 23481707).
Negative control: Mouse liver (PMID: 23481707).
Samples are non-boiled as boiling may cause protein aggregates.
All lanes: Western blot - Anti-NG2 antibody [EPR23976-145] (ab275024) at 1/1000 dilution
Lane 1: Mouse brain tissue lysate at 20 µg
Lane 2: Mouse liver tissue lysate at 20 µg
Lane 3: Mouse pancreas tissue lysate at 40 µg
Lane 4: Rat brain tissue lysate at 40 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 251 kDa
Observed band size: 280 kDa, 330 kDa
Flow Cytometry - Anti-NG2 antibody [EPR23976-145] (ab275024)
Flow cytometric analysis of LADMAC (Mouse bone marrow monocyte macrophage) cells labelling NG2 with ab275024 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
Immunocytochemistry/ Immunofluorescence - Anti-NG2 antibody [EPR23976-145] (ab275024)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized LADMAC cells labelling NG2 with ab275024 at 1/50 (9.34 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in LADMAC cell line. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Western blot - Anti-NG2 antibody [EPR23976-145] (ab275024)
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lane 1: 26 seconds; Lane 2: 59 seconds; Lane 3: 125 seconds.
The band of 330KDa represents the intact NG2 proteoglycan modified by chondroitin sulfate, the band of 280KDa represents NG2 core protein.
The molecular weight observed is consistent with what has been described in the literature (PMID: 20455858, 16625365, 23481707).
Samples are non-boiled as boiling may cause protein aggregates.
All lanes: Western blot - Anti-NG2 antibody [EPR23976-145] (ab275024) at 1/1000 dilution
Lane 1: A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg
Lane 2: SK-MEL-28 (human malignant melanoma) whole cell lysate at 20 µg
Lane 3: Human pancreas tissue lysate at 40 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 251 kDa
Observed band size: 280 kDa, 330 kDa
Immunocytochemistry/ Immunofluorescence - Anti-NG2 antibody [EPR23976-145] (ab275024)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized rat primary neural/glia cell cells labelling NG2 with ab275024 at 1/100 (4.67 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in rat primary glia cells.Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain MAP2 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).
-ve control 1: ab275024 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Flow Cytometry - Anti-NG2 antibody [EPR23976-145] (ab275024)
Flow cytometry overlay histogram showing left A-375 positive cells and right negative MCF7 stained with ab275024 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interactionfollowed by the antibody (ab275024) (1x 106 in 100μl at 5.0 μg/ml (1/402)) for 30min on ice.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.