Anti-Botulinum neurotoxin type A2/BOTOX antibody [EPR28762-552]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Botulinum neurotoxin type A2/BOTOX antibody [EPR28762-552] (AB325604)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T cells transfected with an empty expression vector containing a myc-His-tag®, HEK-293T cells transfected with a BXA1 expression vector containing a myc-His-tag® and HEK-293T cells transfected with a BXA2 expression vector containing a myc-His-tag® cells labelling BOTOX with ab325604 at 1/1000 (0.5 ug/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in HEK-293T cells transfected with a BXA2 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 Myc tag at 1/100 dilution (Magenta).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Botulinum neurotoxin type A2/BOTOX antibody [EPR28762-552] (AB325604)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a Bacteria BXA2 expression vector containing a His, Myc tag, (B) HEK-293T transfected with a Bacteria BXA1 expression vector containing a His, Myc tag and (C) HEK-293T transfected with an empty expression vector containing a His tag tissue labeling BOTOX with ab325604 at 1/1000 (0.5 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a Bacteria BXA2 expression vector containing a His, Myc tag; no staining on both (B) HEK-293T transfected with a Bacteria BXA1 expression vector containing a His, Myc tag and (C) HEK-293T transfected with an empty expression vector containing a 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

• I-ELISA

Lab

Indirect ELISA - Anti-Botulinum neurotoxin type A2/BOTOX antibody [EPR28762-552] (AB325604)

Indirect ELISA analysis of ab325604 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Botulinum neurotoxin type A2/BOTOX.

Antigen concentration : 1000 ng/ml

• WB

Lab

Western blot - Anti-Botulinum neurotoxin type A2/BOTOX antibody [EPR28762-552] (AB325604)

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

This antibody does not cross-react with Bacteria Botulinum neurotoxin type A1 by western blot.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000) (75-150KDa).

All lanes:

Western blot - Anti-Botulinum neurotoxin type A2/BOTOX antibody [EPR28762-552] (ab325604) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 2:

293T transfected with a Bacteria Botulinum neurotoxin type A2/BOTOX expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with a Bacteria Botulinum neurotoxin type A1 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 75-150 kDa,36 kDa

false

Exposure time: 15s