Anti-Strep-tag II antibody [EPR28119-43]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Strep-tag II antibody [EPR28119-43] (AB307676)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Strep-tag II with ab307676 at 1/5000 (0.103 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. Negative control : No staining on human colon. The section was incubated with ab307676 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 LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. 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-Strep-tag II antibody [EPR28119-43] (AB307676)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling Strep-tag II with ab307676 at 1/5000 (0.103 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. Positive staining on (A) HEK-293T cells transfected with a gfp-strep II (WSHPQFEK) expression vector containing a myc-tag. No staining on (B) HEK-293T cells transfected with empty vector containing a myc-tag. The section was incubated with ab307676 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 LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. 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-Strep-tag II antibody [EPR28119-43] (AB307676)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) cells labelling Strep-tag II with ab307676 at 1/100 (5.14 ug/ml) dilution, followed by ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preabsorbed (ab150088) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing positive staining in HEK-293T cells transfected with a Strep-tag II expression vector containing a myc-tag. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 0.38ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preabsorbed (ab150088) at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Strep-tag II antibody [EPR28119-43] (AB307676)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (human embryonic kidney) cells transfected with a Strep-tag II expression vector containing a GFP tag cells labelling Strep-tag II with ab307676 at 1/5000 dilution (0.01ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-Strep-tag II antibody [EPR28119-43] (AB307676)

Strep-tag II was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) cells transfected with a Strep-tag II expression vector containing a GFP tag whole cell lysate 10 ug with ab307676 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307676 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : 293T (human embryonic kidney epithelial cell) cells transfected with a Strep-tag II expression vector containing a GFP tag whole cell lysate 10 ug
Lane 2 : ab307676 IP in 293T cells transfected with a Strep-tag II expression vector containing a GFP tag whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307676 in 293Tcells transfected with a Strep-tag II expression vector containing a GFP tag whole cell lysate

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

Exposure time : 3 seconds

All lanes:

Immunoprecipitation - Anti-Strep-tag II antibody [EPR28119-43] (ab307676) at 1/1000 dilution

Secondary

All lanes:

Immunoprecipitation at 1/5000 dilution

Observed band size: 30 kDa

false

Exposure time: 3s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Strep-tag II antibody [EPR28119-43] (AB307676)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Strep-tag II with ab307676 at 1/5000 (0.103 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. Negative control : No staining on rat colon. The section was incubated with ab307676 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 LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Strep-tag II antibody [EPR28119-43] (AB307676)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Strep-tag II with ab307676 at 1/5000 (0.103 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. Negative control : No staining on mouse colon. The section was incubated with ab307676 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 LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-Strep-tag II antibody [EPR28119-43] (AB307676)

Buffer and diluting concentration : 5% NFDM/TBST In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Primary dilution of 1/1000 and a secondary dilution, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051), of 1/100,000 Exposure time : 3 seconds

All lanes:

Western blot - Anti-Strep-tag II antibody [EPR28119-43] (ab307676) at 1/1000 dilution

Lane 1:

293T cells transfected with a human Strep-tag II expression vector containing a GFP tag whole cell lysate at 20 µg

Lane 2:

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

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 30 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Strep-tag II antibody [EPR28119-43] (AB307676)

Buffer and diluting concentration : 5% NFDM/TBST Primary dilution of 1/1000 and a secondary dilution, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051), of 1/100,000 Exposure time : 1 second

Lane 1:

Western blot - Anti-Strep-tag II antibody [EPR28119-43] (ab307676) at 1/1000 dilution

Lane 1:

Western blot - Anti-Strep-tag II antibody [EPR28119-43] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/strep-tag-ii-antibody-epr28119-43-bsa-and-azide-free-ab307677'>ab307677</a>)

All lanes:

Strep-tagged standard marker (mixture of Strep-tagged recombinant proteins,10-250 kDa)

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 10 kDa,15 kDa,20 kDa,25 kDa,37 kDa,50 kDa,75 kDa,100 kDa,150 kDa,250 kDa

false

Exposure time: 1s