Anti-NG2 antibody [EPR22410-145] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NG2 antibody [EPR22410-145] - BSA and Azide free (AB256351)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling NG2 with ab255811 at 1/100 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : No staining in human colon. (PMID : 25197555). The section was incubated with ab255811 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255811).

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-NG2 antibody [EPR22410-145] - BSA and Azide free (AB256351)

Flow cytometric analysis of SK-BR-3 (Human breast adenocarcinoma epithelial cell, Left panel) / A375 (Human malignant melanoma epithelial cell, Right panel) labeling NG2 with ab255811 at 1/600 (red) compared with a Rabbit monoclonal IgG (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. Negative control : SK-BR-3 (PMID : 20852124). Gated on viable cells.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255811).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NG2 antibody [EPR22410-145] - BSA and Azide free (AB256351)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A375 (human malignant melanoma epithelial cell) cells labeling NG2 with ab255811 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in A375 cells. Negative control : SK-BR-3 (PMID : 20852124). The nuclear counterstain is DAPI (blue). Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red). The negative control is the secondary antibody only.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255811).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NG2 antibody [EPR22410-145] - BSA and Azide free (AB256351)

Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling NG2 with ab255811 at 1/100 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in human melanoma (PMID : 25197555). The section was incubated with ab255811 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255811).

• IP

Supplier Data

Immunoprecipitation - Anti-NG2 antibody [EPR22410-145] - BSA and Azide free (AB256351)

NG2 was immunoprecipitated from 0.35 mg A375 (Human malignant melanoma epithelial cell) whole cell lysate with ab255811 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab255811 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/1000 dilution.

Lane 1 : A375 whole cell lysate 10 μg (Input).
Lane 2 : ab255811 IP in A375 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab255811 in A375 whole cell lysate.

The molecular weight observed is consistent with what has been described in the literature (PMID : 9477982, 8790396). A truncated form is also detected (190kDa, PMID : 25387269).

Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 1 second.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255811).

All lanes:

Immunoprecipitation - Anti-NG2 antibody [EPR22410-145] (<a href='/en-us/products/primary-antibodies/ng2-antibody-epr22410-145-ab255811'>ab255811</a>)

Predicted band size: 251 kDa

Observed band size: 190 kDa,330 kDa,600 kDa

false