Anti-Respiratory Syncytial Virus M2-1 Protein antibody [EPR28649-58]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Respiratory Syncytial Virus M2-1 Protein antibody [EPR28649-58] (AB323319)

Immunohistochemical analysis of paraffin-embedded (A) 293T cells transfected with a Human respiratory syncytial virus A M2-1 protein expression vector containing a his-tag cell pellet. (B) 293T cells transfected with a Human respiratory syncytial virus B M2-1 protein expression vector containing a his-tag cell pellet. (C) 293T transfected with an empty vector containing a his-tag cell pellet. tissue labeling Respiratory Syncytial Virus M2-1 Protein  with ab323319 at 1/10000 (0.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) 293T transfected with a HRSVA M2-1 expression vector containing a his-tag cell pellet and (B) 293T transfected with a HRSVB M2-1 expression vector containing a his-tag cell pellet. No staining in (C) 293T transfected with an empty vector containing a his-tag cell pellet.
The section was incubated with ab323319 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

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Immunocytochemistry/ Immunofluorescence - Anti-Respiratory Syncytial Virus M2-1 Protein antibody [EPR28649-58] (AB323319)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Respiratory Syncytial Virus M2-1 Protein  with ab323319 at 1/4000 (0.129 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing positive staining in 293T cells transfected with a Human respiratory syncytial virus A (HRSVA) and Human respiratory syncytial virus B (HRSVB) M2-1 protein expression vectors containing a myc-His-tag® (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml 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 2 ug/ml dilution.

• I-ELISA

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Indirect ELISA - Anti-Respiratory Syncytial Virus M2-1 Protein antibody [EPR28649-58] (AB323319)

Indirect ELISA analysis of ab323319 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 : Respiratory Syncytial Virus M2-1 protein, Respiratory Syncytial Virus M2-1 peptide.

Antigen concentration : 1000 ng/ml

• WB

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Western blot - Anti-Respiratory Syncytial Virus M2-1 Protein antibody [EPR28649-58] (AB323319)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Respiratory Syncytial Virus M2-1 Protein antibody [EPR28649-58] (ab323319) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Human respiratory syncytial virus A M2-1 protein expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a Human respiratory syncytial virus B M2-1 protein 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='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 22 kDa,36 kDa

false

Exposure time: 3s