Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric)

8 product images

• 

  Immunoprecipitation - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294)

  RILPL1 was immunoprecipitated from 0.35 mg rat brain tissue lysate 10 µg with ab307294 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307294 at dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Rat brain tissue lysate 10 µg

  Lane 2: ab307294 at 1/30 IP in rat brain tissue lysate 10 µg

  Lane 3: Mouse IgG2a Isotype control (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) instead of ab307294 in rat brain tissue lysate

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

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

  All lanes: Immunoprecipitation - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294) at 1/1000 dilution

  Lane 1: Rat brain tissue lysate at 10 µg

  Lane 2: ab307294 at 1/30 IP in rat brain tissue lysate at 10 µg

  Lane 3: Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413 in rat brain tissue lysate at 10 µg

  Secondary

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

  Developed using the ECL technique.

  Observed band size: 47 kDa

  Exposure time: 3min

• 

  Immunoprecipitation - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294)

  RILPL1 was immunoprecipitated from 0.35 mg mouse brain tissue lysate 10 µg with ab307294 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307294 at dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Mouse brain tissue lysate 10 µg

  Lane 2: ab307294 at 1/30 IP in mouse brain tissue lysate 10 µg

  Lane 3: Mouse IgG2a Isotype control (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) instead of ab307294 in mouse brain tissue lysate

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

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

  All lanes: Immunoprecipitation - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294) at 1/1000 dilution

  Lane 1: Mouse brain tissue lysate at 10 µg

  Lanes 1 - 2: ab307294 at 1/30 IP in mouse brain tissue lysate at 10 µg

  Lane 3: Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413 in mouse brain tissue lysate (isotype control)

  Secondary

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

  Developed using the ECL technique.

  Observed band size: 47 kDa

  Exposure time: 3min

  RILPL1 was immunoprecipitated from 0.35 mg mouse brain tissue lysate 10 µg with ab307294 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307294 at dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Mouse brain tissue lysate 10 µg

  Lane 2: ab307294 at 1/30 IP in mouse brain tissue lysate 10 µg

  Lane 3: Mouse IgG2a Isotype control (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) instead of ab307294 in mouse brain tissue lysate

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

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

• 

  Immunoprecipitation - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294)

  RILPL1 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell) whole cell lysate 10 µg with ab307294 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307294 at dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: A549 whole cell lysate 10 µg

  Lane 2: ab307294 at 1/30 IP in A549 whole cell lysate 10 µg

  Lane 3: Mouse IgG2a Isotype control (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) instead of ab307294 in A549 whole cell lysate

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

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

  All lanes: Immunoprecipitation - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294) at 1/1000 dilution

  Lane 1: A549 (human lung carcinoma epithelial cell) at 10 µg

  Lane 2: ab307294 at 1/30 IP in A549 (human lung carcinoma epithelial cell) at 10 µg

  Lane 3: Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413 in A549 whole cell lysate

  Secondary

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

  Developed using the ECL technique.

  Observed band size: 47 kDa

  Exposure time: 3min

• 

  Western blot - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294)

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

  Low expression: U937 (Human Protein Atlas).

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

  All lanes: Western blot - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294) at 1/1000 dilution

  Lane 1: SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

  Lane 2: HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

  Lane 3: U937 (human histiocytic lymphoma monocyte), whole cell lysate at 20 µg

  Lane 4: Human cerebellum tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG Fc (HRP) preadsorbed (Goat Anti-Mouse IgG Fc (HRP) preadsorbed ab98717) at 1/20000 dilution

  Developed using the ECL technique.

  Observed band size: 47 kDa

  Exposure time: 81s

• 

  Western blot - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294)

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

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

  Exposure time:

  Lane 1-4: 180 seconds

  Lanes 5-7: 125 seconds

  All lanes: Western blot - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a (Chimeric) (ab307294) at 1/1000 dilution

  Lane 1: C6 (rat glial tumor cell), whole cell lysate at 10 µg

  Lane 2: PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

  Lane 3: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

  Lane 4: Rat brain tissue lysate at 10 µg

  Lane 5: Mouse brain tissue lysate at 10 µg

  Lane 6: Mouse heart tissue lysate at 10 µg

  Lane 7: Mouse spleen tissue lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG Fc (HRP) preadsorbed (Goat Anti-Mouse IgG Fc (HRP) preadsorbed ab98717) at 1/20000 dilution

  Developed using the ECL technique.

  Observed band size: 47 kDa

• 

  Western blot - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a - Chimeric (ab307294)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  In Western blot, ab307294 was shown to bind specifically to RILPL1. A band was observed at 47 kDa in wild-type A549 cell lysates with whereas no signal observed at this size in RILPL1 knockout cell line.
  
  Wild-type and RILPL1 knockout A549 cell lysates were kindly provided by Dr. Dario Alessi, University of Dundee.
  
  Exposure time: 3 minutes.

  All lanes: Western blot - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a - Chimeric (ab307294) at 1/1000 dilution

  Lane 1: Wild-type A549 (human lung carcinoma epithelial cell), whole cell lysate 20 μg at 20 µg

  Lane 2: RILPL1 knockout A549 (human lung carcinoma epithelial cell), whole cell lysate 20 μg at 20 µg

  Lane 3: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 20 μg at 20 µg

  Lane 4: MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate 20 μg at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG Fc (HRP) preadsorbed (Goat Anti-Mouse IgG Fc (HRP) preadsorbed ab98717) at 1/20000 dilution

  Observed band size: 47 kDa

  Exposure time: 3min

• 

  Immunocytochemistry/ Immunofluorescence - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a - Chimeric (ab307294)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labeling RILPL1 with ab307294 at 1/500 dilution, followed by Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed ab98712 Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green).
  Confocal image showing centrosome and cytoplasmic staining in SH-SY5Y cell line.
  
  Low expression: U-937.
  Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (10µg/ml) followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/400 dilution (4 µg/ml) (Red). The nuclear counterstain was DAPI (Blue).
  
  -ve control 1: ab307294 at a 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 at a 1/500 dilution.
  
  -ve control 2: Anti-beta Tubulin antibody [EPR16774] ab179513 at a 1/200 dilution followed by Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed ab98712 at a 1/1000 dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-RILPL1 antibody [MJF-R41-21] - Mouse IgG2a - Chimeric (ab307294)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labeling RILPL1 with ab307294 at 1/500 dilution, followed by Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed ab98712 Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green).
  Confocal image showing centrosome staining in Neuro-2a cell line.
  Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (10µg/ml) followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/400 dilution (4 µg/ml) (Red). The nuclear counterstain was DAPI (Blue).
  
  -ve control 1: ab307294 at a 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 at a 1/500 dilution.
  
  -ve control 2: Anti-beta Tubulin antibody [EPR16774] ab179513 at a 1/200 dilution followed by Goat Anti-Mouse IgG Fc (DyLight® 488) preadsorbed ab98712 at a 1/1000 dilution.