Anti-Streptavidin antibody [EPR28602-42]

• Flow Cyt

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Flow Cytometry - Anti-Streptavidin antibody [EPR28602-42] (AB319103)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling Streptavidin with ab319103 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Cells were co-stained with biotin-anti human CD4 and streptavidin conjugated to PE.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Streptavidin antibody [EPR28602-42] (AB319103)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a Streptavidin expression vector containing a his tag. (B) HEK-293T transfected with empty vector containing a his tag tissue labeling Streptavidin with ab319103 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a Streptavidin expression vector containing a his tag. No staining on (B) HEK-293T transfected with empty vector containing a his tag.
The section was incubated with ab319103 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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Streptavidin antibody [EPR28602-42] (AB319103)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Streptavidin with ab319103 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse liver.
The section was incubated with ab319103 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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Streptavidin antibody [EPR28602-42] (AB319103)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Streptavidin with ab319103 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat liver.
The section was incubated with ab319103 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

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Immunocytochemistry/ Immunofluorescence - Anti-Streptavidin antibody [EPR28602-42] (AB319103)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Streptavidin with ab319103 at 1/4000 (0.126 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 293T cells transfected with a Streptavidin expression vector 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).

Myc-Tag Mouse mAb (Alexa Fluor® 647) 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.

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Streptavidin antibody [EPR28602-42] (AB319103)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T (human embryonic kidney epithelial cell) transfected with a Streptavidin expression vector containing a myc-His-tag® (Middle) / 293T transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling Streptavidin with ab319103 at 1/5000 dilution (0.1ug) / Middle and right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti myc tag conjugated to Alexa Fluor®647.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Streptavidin antibody [EPR28602-42] (AB319103)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Streptavidin with ab319103 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human liver.
The section was incubated with ab319103 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

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Indirect ELISA - Anti-Streptavidin antibody [EPR28602-42] (AB319103)

Indirect ELISA analysis of ab319103 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Streptavidin.

Antigen concentration : 1000 ng/ml

• WB

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Western blot - Anti-Streptavidin antibody [EPR28602-42] (AB319103)

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-Streptavidin antibody [EPR28602-42] (ab319103) at 1/1000 dilution

Lane 1:

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

Lane 2:

293T transfected with a Streptavidin 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 1/20000 dilution

Observed band size: 21 kDa,36 kDa

false

Exposure time: 180s