Anti-4 Hydroxynonenal antibody [HNEJ-2]

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-4 Hydroxynonenal antibody [HNEJ-2] (AB48506)

Elevated brain oxidative damage in ischemic 5xFAD mice

Formalin-fixed, paraffin-embedded mouse brain tissue (from non-trasngenic or 5xFAD transgenic animals) stained for 4 Hydroxynonenal using ab48506 at in immunohistochemical analysis.

Scale bar = 20μm. (A2) and (B2) are representative images of 4-HNE staining (Red) in brain cortex and CA1 region, respectively.

Lu, L. et al PLoS One. 2015 Dec 3;10(12):e0144068. doi: 10.1371/journal.pone.0144068. eCollection 2015. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• IHC-P

AbReview16917****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-4 Hydroxynonenal antibody [HNEJ-2] (AB48506)

ab48506 at a 1/25 dilution staining 4-Hydroxy-2-Nonenal in mouse heart tissue sections by Immunohistochemistry (paraffin embedded) incubated for 15 hours at +4°C. Fixed with formaldehyde, heat mediated antigen retrieval step performed using citrate buffer. Blocked using 5% serum for 20 minutes at 20°C. Secondary used undiluted polyclonal Goat anti-mouse IgG conjugated to Alexa Fluor 594.

This image was kindly supplied by Dr Jinqing Li by Abreview

• IHC-FoFr

AbReview33645****

Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-4 Hydroxynonenal antibody [HNEJ-2] (AB48506)

ab15463 staining 4-Hydroxynonenal in the Mouse melanoma cancer tissue sections by IHC-FoFr (PFA perfusion fixed frozen Sections). Tissue samples were fixed with 4% Formalin, Antigen retrieval carried out heat mediation using anautoclave for 10 minutes, permeabilized using 0.1% SDS and blocked with 1% BSAfor 15 minutes at 25°C. The sample was incubated with primary antibody (1/25 in PBS + 1% BSA) at 4°C for 12 hours.An Alexa Fluor^®488-conjugated Goat anti-mouse IgG(H+L) monoclonal(1/100) was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Seontae Kim

• WB

Lab

Western blot - Anti-4 Hydroxynonenal antibody [HNEJ-2] (AB48506)

Western blot : Anti-4-HNE antibody [HNEJ-2] (ab48506) staining at 1/1000 dilution, shown in black. In Western blot, ab48506 was shown to bind specifically to 4-HNE. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 3 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Mouse (H+L) 5000 dilution.

All lanes:

Western blot - Anti-4 Hydroxynonenal antibody [HNEJ-2] (ab48506) at 1/1000 dilution

Lane 1:

BSA cell lysate at 0.5 µg

Lane 2:

BSA cell lysate at 1 µg

Lane 3:

4-Hydroxynonenal (BSA) cell lysate at 0.5 µg

Lane 4:

4-Hydroxynonenal (BSA) cell lysate at 1 µg

Observed band size: 66 kDa

true

• WB

Lab

Western blot - Anti-4 Hydroxynonenal antibody [HNEJ-2] (AB48506)

Western blot : Anti-4-HNE antibody (ab46545) staining at 1/1000 dilution, shown in black. In Western blot, ab46545 binds to 4-HNE but shows some non-specific binding to BSA. We recommend ab48506 for Western blot of 4-HNE. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 3 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) at 1/50000 dilution.

Lanes 1 - 4:

Western blot - Anti-4 Hydroxynonenal antibody (<a href='/en-us/products/primary-antibodies/4-hydroxynonenal-antibody-ab46545'>ab46545</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-4 Hydroxynonenal antibody [HNEJ-2] (ab48506) at 1/1000 dilution

Lane 1:

BSA cell lysate at 0.5 µg

Lane 2:

BSA cell lysate at 1 µg

Lane 3:

4-Hydroxynonenal (BSA) cell lysate at 0.5 µg

Lane 4:

4-Hydroxynonenal (BSA) cell lysate at 1 µg

Observed band size: 66 kDa

true