Anti-Biotin antibody [EPR28239-85]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Biotin antibody [EPR28239-85] (AB324266)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Biotin with ab324266 at 1/1000 (0.525 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 1000 2ug/ml dilution (Green).

Confocal image showing positive staining in HeLa cells (shown in green) pre-incubated with 0.5 ug/ml biotinylated Anti-Vimentin antibody [EPR3776] – Chicken IgY (Chimeric) (ab316214) at 37℃ for 1 hour (Panel A). In contrast, no positive staining is detected when the cells are incubated only with the Anti-Biotin antibody (ab324266) under the same conditions(Panel B). 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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ab195889 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/ab150081 1000 2ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Biotin antibody [EPR28239-85] (AB324266)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Biotin with ab324266 at 1/2000 (0.263 ug/ml) dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880).

Positive staining in human pancreas tissue pre-incubated at 37°C for 1 hour with biotinylated Anti-Vimentin antibody [EPR3776] – Chicken IgY (Chimeric) (ab316214), followed by overnight incubation at 4°C with Rabbit monoclonal Anti-Biotin antibody [EPR28239-85] (ab324266) (Image A). In contrast, no positive staining is detected when the tissue is incubated only with the Anti-Biotin antibody (ab324266) under the same conditions (Image B).

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880).

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-Biotin antibody [EPR28239-85] (AB324266)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling Biotin with ab324266 at 1/2000 (0.263 ug/ml) dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880).

Positive staining in human breast tissue pre-incubated at 37°C for 1 hour with biotinylated Anti-Vimentin antibody [EPR3776] – Chicken IgY (Chimeric) (ab316214), followed by overnight incubation at 4°C with Rabbit monoclonal Anti-Biotin antibody [EPR28239-85] (ab324266) (Image A). In contrast, no positive staining is detected when the tissue is incubated only with the Anti-Biotin antibody (ab324266) under the same conditions (Image B).

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880).

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-Biotin antibody [EPR28239-85] (AB324266)

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

Western Blot analysis of specific lysates using Anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) (ab316214) biotinylated, followed by Rabbit monoclonal antibody [EPR28239-85] to Biotin (ab324266) and Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

All lanes:

Western blot - Anti-Biotin antibody [EPR28239-85] (ab324266) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate pre-incubated with anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) (<a href='/en-us/products/primary-antibodies/vimentin-antibody-epr3776-chicken-igy-chimeric-ab316214'>ab316214</a>) biotinylated at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate pre-incubated with anti-Vimentin antibody [EPR3776] - Chicken IgY (Chimeric) (<a href='/en-us/products/primary-antibodies/vimentin-antibody-epr3776-chicken-igy-chimeric-ab316214'>ab316214</a>) biotinylated 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: 53 kDa

false

Exposure time: 92s

• I-ELISA

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Indirect ELISA - Anti-Biotin antibody [EPR28239-85] (AB324266)

Indirect ELISA analysis of ab324266 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 : Biotin-crosslink-BSA, BSA.

Antigen concentration : 1000 ng/ml