MA1-90928

antibody from Invitrogen Antibodies

Targeting: CSPG4 CSPG4A, HMW-MAA, MCSP, MCSPG, MEL-CSPG, MSK16, NG2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Recommended by provider)

Flow cytometry (Recommended by provider)

Antibody Data

Product number

MA1-90928

Provider

Invitrogen Antibodies

Product name

NG2 Monoclonal Antibody (LHM 2)

Antibody type

Monoclonal

Antigen

Other

Description

MA1-90928 detects MCSP from human samples.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

LHM 2

Vial size

100 µg

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Monitoring the systemic human memory B cell compartment of melanoma patients for anti-tumor IgG antibodies.

Gilbert AE, Karagiannis P, Dodev T, Koers A, Lacy K, Josephs DH, Takhar P, Geh JL, Healy C, Harries M, Acland KM, Rudman SM, Beavil RL, Blower PJ, Beavil AJ, Gould HJ, Spicer J, Nestle FO, Karagiannis SN
PloS one 2011 Apr 29;6(4):e19330

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using NG2 Monoclonal Antibody (LHM 2) (Product # MA1-90928) and a band at ~250 kDa corresponding to NG2 was observed only in A-375 when compared to other cell lines which are reported negative. Whole cell extracts (30 µg lysate) of A-375 (Lane 1), K-562 (Lane 2), U-2 OS (Lane 3), HeLa (Lane 4), MCF-7 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of NG2 was performed using 70% confluent log phase A-375 cells. The cells were fixed with 4% Paraformaldehyde for 10 Minutes, permeabilized with 0.1% Triton™ X-100 for 10 Minutes, and blocked with 2% BSA for 10 Minutes at room temperature. The cells were labeled with NG2 Monoclonal Antibody (LHM 2) (Product # MA1-90928) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175, 1:2,000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization. Panel e represents HeLa cells with no expression for NG2. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.