Anti-Ferredoxin, apicoplast antibody [EPR31001-512R]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ferredoxin, apicoplast antibody [EPR31001-512R] (AB325924)

Immunohistochemical analysis of paraffin-embedded A : HEK-293T (human epithelial cell line from embryonic kidney) transfected with a Plasmodium falciparum Ferredoxin expression vector containing a Myc-His tag B : HEK-293T transfected with empty vector containing a Myc-His tag tissue labeling Ferredoxin, apicoplast with ab325924 at 1/5000 (0.1 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in A : HEK-293T transfected with a Plasmodium falciparum Ferredoxin expression vector containing a Myc-His tag, and no signal in B : HEK-293T transfected with empty vector containing a Myc-His tag.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Ferredoxin, apicoplast antibody [EPR31001-512R] (AB325924)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells transfected with empty vector containing a myc-His-tag®. 293T cells transfected with a Plasmodium falciparum Ferredoxin expression vector containing a myc-His-tag®. cells labelling Ferredoxin, apicoplast with ab325924 at 1/500 (1.0 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in 293T cells transfected with a Plasmodium falciparum Ferredoxin expression vector containing a myc-His-tag® (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• WB

Lab

Western blot - Anti-Ferredoxin, apicoplast antibody [EPR31001-512R] (AB325924)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa);Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(20-60KDa).

All lanes:

Western blot - Anti-Ferredoxin, apicoplast antibody [EPR31001-512R] (ab325924) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Plasmodium falciparum Ferredoxin expression vector containing a his-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 20-60 kDa,36 kDa

false

Exposure time: 10s