Anti-CLEC2D antibody [EPR27404-37]

3 product images

• 

  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)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  Lane 1: Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate
  Lane 2: abab309147 IP in Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate
  Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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 1/30 dilution

  All lanes: Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

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

  Exposure time: 6s

• 

  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 (Anti-GAPDH antibody [EPR16891] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

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

  Exposure time: 92s

• 

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.