Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free (AB220811)

Immunohistochemical staining of paraffin embedded human pancreas with purified ab76310 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free (AB220811)

Overlay histogram showing HeLa cells stained with unpurified ab76310 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76310, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free (AB220811)

Immunohistochemical staining of paraffin-embedded human gastric carcinoma using unpurified ab76310 at 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC

Lab

Immunohistochemistry - Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free (AB220811)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

Immunohistochemical analysis of formalin fixed paraffin embedded human pancreas labelling DUSP6 with ab76310 at a concentration of 0.5 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32 mins.

ab76310 anti-DUSP6 [EPR129Y] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free (AB220811)

Overlay histogram showing NIH-3T3 cells fixed in 4% PFA and stained with purified ab76310 at a dilution of 1 in 200 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

• IP

Unknown

Immunoprecipitation - Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free (AB220811)

ab76310 (purified) at 1/20 immunoprecipitating DUSP6 in 10 µg NIH-3T3 (Lanes 1 and 2, observed at 42 and 44 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBSTThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76310).

All lanes:

Immunoprecipitation - Anti-DUSP6 antibody [EPR129Y] (<a href='/en-us/products/primary-antibodies/dusp6-antibody-epr129y-ab76310'>ab76310</a>)

Predicted band size: 42 kDa

false

• WB

PubMed

Western blot - Anti-DUSP6 antibody [EPR129Y] - BSA and Azide free (AB220811)

Immunoblot analysis of indicated proteins in splenocytes after CD4+ sort, liver homogenate or purified CD4+ T cells from WT, DUSP6+/- or DUSP6-/- mice to confirm knockout or heterozygosity for DUSP6 at protein level.

All lanes:

Western blot - Anti-DUSP6 antibody [EPR129Y] (<a href='/en-us/products/primary-antibodies/dusp6-antibody-epr129y-ab76310'>ab76310</a>)

Predicted band size: 42 kDa

false

Schmidt C et al., Front Immunol, 9, 2386, 2018 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/