Anti-CLEC2D antibody [EPR27404-37]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CLEC2D antibody [EPR27404-37] (AB309147)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labelling CLEC2D with ab309147 at 1/50 (9.88 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 cytoplasmic staining in Raji cell line, no staining was observed in HeLa cell line.Negative control : HeLa (PMID : 26405582).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.

• IP

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Immunoprecipitation - Anti-CLEC2D antibody [EPR27404-37] (AB309147)

CLEC2D was immunoprecipitated from 0.35 mg Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate with ab309147 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309147 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate
Lane 2 : ab309147 IP in Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab309147 in Raji whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-CLEC2D antibody [EPR27404-37] (ab309147) at 1/1000 dilution

All lanes:

Raji (human burkitt's lymphoma b lymphocyte) 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

Observed band size: 33 kDa,22 kDa,18 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-CLEC2D antibody [EPR27404-37] (AB309147)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : HeLa (PMID : 26405582), 293T (PMID : 20843815). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CLEC2D antibody [EPR27404-37] (ab309147) at 1/1000 dilution

Lane 1:

Ramos (human burkitts lymphoma b lymphocyte) whole cell lysate at 20 µg

Lane 2:

Raji (human burkitts lymphoma b lymphocyte) whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate

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: 33 kDa,22 kDa,18 kDa

false

Exposure time: 92s