ET1703-16

NG2 Recombinant Rabbit Monoclonal Antibody [JM10-13]

cat.: ET1703-16

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JM10-13

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 251 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human NG2 aa 2,271-2,314 / 2,322.
Positive control:	A375 cell lysate, SK-MEL-28 cell lysate, MCF7 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, SiHa cell lysate, THP-1 cell lysate, rat brain tissue, mouse brain tissue, SHG-44, human malignant melanoma tissue.
Subcellular location:	Cell membrane, Apical cell membrane, Cell projection, Cell surface.
Recommended Dilutions: WB IHC-P FC	1:1,000-1:2,000 1:50-1:400 1:50-1:100
Uniprot #:	SwissProt: Q6UVK1 Human \| CSPG4 Mouse \| Q00657 Rat
Alternative names:	4732461B14Rik AN2 AN2 proteoglycan Chondroitin sulfate proteoglycan 4 (melanoma-associated) Chondroitin sulfate proteoglycan 4 Chondroitin sulfate proteoglycan NG2 Cspg4 Cspg4 chondroitin sulfate proteoglycan 4 CSPG4_HUMAN HMW-MAA HSN tumor-specific antigen Kiaa4232 MCSP MCSPG MEL-CSPG Melanoma chondroitin sulfate proteoglycan Melanoma-associated chondroitin sulfate proteoglycan MELCSPG MSK16 NG2

Images

Fig1: Western blot analysis of NG2 on different lysates with Rabbit anti-NG2 antibody (ET1703-16) at 1/2,000 dilution.

Lane 1: A375 cell lysate (15 µg/Lane)
Lane 2: SK-MEL-28 cell lysate (15 µg/Lane)
Lane 3: MCF7 cell lysate (negative control) (15 µg/Lane)
Lane 4: Mouse brain tissue lysate (20 µg/Lane)
Lane 5: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 251 kDa
Observed band size: 300 kDa

Exposure time: 2 minutes;

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 (ET1703-16) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of NG2 on different lysates with Rabbit anti-NG2 antibody (ET1703-16) at 1/1,000 dilution.

Lane 1: SiHa cell lysate
Lane 2: THP-1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 251 kDa
Observed band size: 300 kDa

Exposure time: 3 minutes;

6% 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 (ET1703-16) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-NG2 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 ddH₂O and PBS, and then probed with the primary antibody (ET1703-16, 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 mouse brain tissue using anti-NG2 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 ddH₂O and PBS, and then probed with the primary antibody (ET1703-16, 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: Flow cytometric analysis of NG2 was done on SHG-44 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-16, 1/50) (red). 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 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig6: Immunohistochemical analysis of paraffin-embedded human malignant melanoma tissue using anti-NG2 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 1% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1703-16, 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.