Anti-Cholera enterotoxin subunit B antibody [EPR27372-4]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cholera enterotoxin subunit B antibody [EPR27372-4] (AB317738)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a Cholera enterotoxin subunit B expression vector containing a EGFP tag. (B) HEK-293T transfected with empty vector containing a EGFP tag. tissue labeling Cholera enterotoxin subunit B with ab317738 at 1/2000 (0.265 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a Cholera enterotoxin subunit B expression vector containing a EGFP tag, negative staining on (B) HEK-293T transfected with empty vector containing a EGFP tag. The section was incubated with ab317738 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-Cholera enterotoxin subunit B antibody [EPR27372-4] (AB317738)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Cholera enterotoxin subunit B with ab317738 at 1/2000 (0.265 ug/ml) dilution, followed by ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150088) antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing positive staining in 293T cells transfected with a Cholera enterotoxin subunit B expression vector (shown in magenta) containing a EGFP-tag® (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody only control : Secondary antibody is ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150088) at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cholera enterotoxin subunit B antibody [EPR27372-4] (AB317738)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a Cholera enterotoxin subunit B expression vector containing an EGFP tag (Middle) / 293T cells transfected with an empty expression vector containing an EGFP tag (Right) cells labelling Cholera enterotoxin subunit B with ab317738 at 1/5000 dilution (0.01 ug)/Middle and Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-Cholera enterotoxin subunit B antibody [EPR27372-4] (AB317738)

Cholera enterotoxin subunit B was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) transfected with a Cholera enterotoxin subunit B expression vector containing a EGFP tag, whole cell lysate with ab317738 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317738 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) transfected with a Cholera enterotoxin subunit B expression vector containing a EGFP tag, whole cell lysate
Lane 2 : ab317738 IP in 293T (human embryonic kidney epithelial cell) transfected with a Cholera enterotoxin subunit B expression vector containing a EGFP tag, whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317738 in 293T cells transfected with a Cholera enterotoxin subunit B expression vector containing a EGFP tag, whole cell lysate

All lanes:

Immunoprecipitation - Anti-Cholera enterotoxin subunit B antibody [EPR27372-4] (ab317738) at 1/30 dilution

All lanes:

293T (human embryonic kidney epithelial cell) transfected with a Cholera enterotoxin subunit B expression vector containing a EGFP tag, whole cell lysate at 10 µg

Secondary

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Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 10s

• ELISA

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ELISA - Anti-Cholera enterotoxin subunit B antibody [EPR27372-4] (AB317738)

Indirect ELISA analysis of ab317738 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 : Cholera enterotoxin subunit B.

Antigen concentration : 1000 ng/ml

The recombinant protein of Cholera enterotoxin subunit B was made in house.

• WB

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Western blot - Anti-Cholera enterotoxin subunit B antibody [EPR27372-4] (AB317738)

In Western blot, Anti-GFP antibody [E385] (ab32146) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Cholera enterotoxin subunit B antibody [EPR27372-4] (ab317738) at 1/5000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a EGFP tag, whole cell lysate at 2 µg

Lane 2:

293T transfected with a Cholera enterotoxin subunit B expression vector containing a EGFP tag, whole cell lysate 2 at 2 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: 40 kDa

false

Exposure time: 180s