Anti-BASP1 antibody [EPR28065-75]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BASP1 antibody [EPR28065-75] (AB312322)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscl tissue labeling BASP1 with ab312322 at 1/2000 (0.246 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human skeletal muscle.The section was incubated with ab312322 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-BASP1 antibody [EPR28065-75] (AB312322)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling BASP1 with ab312322 at 1/2000 (0.246 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum.The section was incubated with ab312322 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-BASP1 antibody [EPR28065-75] (AB312322)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling BASP1 with ab312322 at 1/2000 (0.246 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil.The section was incubated with ab312322 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-BASP1 antibody [EPR28065-75] (AB312322)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling BASP1 with ab312322 at 1/50 (9.84 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing membranous and weak cytoplasmic staining in HeLa cell line.Negative control : K-562 (PMID : 14701728).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). 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 2ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-BASP1 antibody [EPR28065-75] (AB312322)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast, Left) / HeLa (human cervical adenocarcinoma epithelial cell, Right) cells labelling BASP1 with ab312322 at 1/500 dilution (0.1 ug)/Red (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). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Negative control : K-562 (PMID : 14701728).

• IP

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Immunoprecipitation - Anti-BASP1 antibody [EPR28065-75] (AB312322)

BASP1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab312322 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab312322 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate Lane 2 : ab312322 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab312322 in HeLa whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-BASP1 antibody [EPR28065-75] (ab312322) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

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: 180s

• WB

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Western blot - Anti-BASP1 antibody [EPR28065-75] (AB312322)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : skeletal muscle, liver (9749536, 14701728). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 15 seconds.

All lanes:

Western blot - Anti-BASP1 antibody [EPR28065-75] (ab312322) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 55 kDa

false

Exposure time: 15s

• WB

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Western blot - Anti-BASP1 antibody [EPR28065-75] (AB312322)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : K-562 (PMID : 14701728). The molecular weight observed is consistent with what has been described in the literature (PMID : 33692421; 8390468). In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution. Exposure time : 103 seconds.

All lanes:

Western blot - Anti-BASP1 antibody [EPR28065-75] (ab312322) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA 1 specifically targeti BASP1 whole cell lysate at 20 µg

Lane 3:

HeLa transfected with siRNA 2 specifically targeti BASP1 whole cell lysate at 20 µg

Lane 4:

K-562 (human chronic myelogenous leukemia lymphoblast) 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: 55 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-BASP1 antibody [EPR28065-75] (AB312322)

Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody does not cross-react with mouse BASP1. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 59 seconds.

All lanes:

Western blot - Anti-BASP1 antibody [EPR28065-75] (ab312322) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containi a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human BASP1 expression vector containi a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a mouse BASP1 expression vector containi 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 1/100000 dilution

Observed band size: 55 kDa

false

Exposure time: 59s