Anti-RAGE antibody [RM1078]

16 product images

• 

  Western blot - Anti-RAGE antibody [RM1078] (ab314773)

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

  Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as loading control at a 1/1000000 dilution.

  All lanes: Western blot - Anti-RAGE antibody [RM1078] (ab314773) at 1/1000 dilution

  Lane 1: Mouse brain tissue lysate at 20 µg

  Lane 2: Mouse brain tissue lysate at 40 µg

  Lane 3: Mouse kidney tissue lysate at 20 µg

  Lane 4: Mouse Kidney tissue lysate at 40 µg

  Lane 5: Mouse spinal cord tissue lysate at 20 µg

  Lane 6: Mouse spinal cord tissue lysate at 40 µg

  Lane 7: HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

  Lane 8: Blank

  Lane 9: Mouse lung tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 42 kDa

  Observed band size: 45 kDa, 55 kDa

  Exposure time: 180s

• 

  Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat lung (fresh) tissue labeling RAGE with ab314773 at 1/100 (5.17 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 at 1/1000 2 ug/mL dilution (Green).
  Confocal image showing positive staining on rat lung. The nuclear counterstain was DAPI (Blue). The section was incubated with ab314773 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).
  Secondary antibody 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 2 ug/mL dilution.

• 

  Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver (fresh) tissue labeling RAGE with ab314773 at 1/100 (5.17 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 at 1/1000 2 ug/mL dilution (Green).
  Negative control: confocal image showing no staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab314773 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).
  Secondary antibody 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 2 ug/mL dilution.

• 

  Western blot - Anti-RAGE antibody [RM1078] (ab314773)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  Low expression: liver, spleen, kidney and skeletal muscle (PMID:16315007; PMID:18245812). The expression profile and molecular mass observed is consistent with what has been described in the literature (PMID:16315007; PMID:18355449; PMID:18245812). In western blot, anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  All lanes: Western blot - Anti-RAGE antibody [RM1078] (ab314773) at 1/1000 dilution

  Lane 1: Mouse lung tissue lysate at 20 µg

  Lane 2: Mouse spleen tissue lysate at 20 µg

  Lane 3: Mouse liver tissue lysate at 20 µg

  Lane 4: Mouse kidney tissue lysate at 20 µg

  Lane 5: Rat lung tissue lysate at 20 µg

  Lane 6: Rat spleen tissue lysate at 20 µg

  Lane 7: Rat liver tissue lysate at 20 µg

  Lane 8: Rat skeletal muslce tissue lysate at 20 µg

  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: 45 kDa, 55 kDa

  Exposure time: 15s

• 

  Western blot - Anti-RAGE antibody [RM1078] (ab314773)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  Low expression: liver and skeletal muscle (PMID:16315007; PMID:18245812). The expression profile and molecular mass observed is consistent with what has been described in the literature (PMID:16315007; PMID:18355449; PMID:18245812). In western blot, anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  All lanes: Western blot - Anti-RAGE antibody [RM1078] (ab314773) at 1/1000 dilution

  Lane 1: Human lung 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: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 45 kDa, 55 kDa

  Exposure time: 26s

• 

  Immunocytochemistry/ Immunofluorescence - Anti-RAGE antibody [RM1078] (ab314773)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney) transfected with myc-tagged AGER expression vector cells labelling RAGE with ab314773 at 1/500 (1.034 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 membranous or cytoplasmic staining in 293T cells transfected with AGER protein expression vector containing a myc tag, no staining on 293T transfected with an empty vector containing a myc-tag.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
  Alexa Fluor® 594 Anti-Myc tag antibody [9E10] ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling RAGE with ab314773 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Negative control: no staining on rat liver. The section was incubated with ab314773 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling RAGE with ab314773 at 1/1000 (0.517 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Negative control: no staining on mouse liver. The section was incubated with ab314773 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of paraffin-embedded human liver tissue labeling RAGE with ab314773 at 1/1000 (0.517 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Negative control: no staining on human liver. The section was incubated with ab314773 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of paraffin-embedded rat lung tissue labeling RAGE with ab314773 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Positive staining on rat lung. The section was incubated with ab314773 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling RAGE with ab314773 at 1/1000 (0.517 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Positive staining on mouse lung. The section was incubated with ab314773 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of paraffin-embedded human lung tissue labeling RAGE with ab314773 at 1/1000 (0.517 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
  Positive staining on human lung. The section was incubated with ab314773 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• 

  Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver (fresh) tissue labeling RAGE with ab314773 at 1/100 (5.17 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 at 1/1000 2 ug/mL dilution (Green).
  Negative control: confocal image showing no staining on rat liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab314773 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).
  Secondary antibody 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 2 ug/mL dilution.

• 

  Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [RM1078] (ab314773)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse lung (fresh) tissue labeling RAGE with ab314773 at 1/100 (5.17 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 at 1/1000 2 ug/mL dilution (Green).
  Confocal image showing positive staining on mouse lung. The nuclear counterstain was DAPI (Blue). The section was incubated with ab314773 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).
  Secondary antibody 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 2 ug/mL dilution.

• 

  Immunoprecipitation - Anti-RAGE antibody [RM1078] (ab314773)

  RAGE was immunoprecipitated from 0.35 mg Mouse lung tissue lysate with ab314773 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314773 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: Mouse lung tissue lysate
  Lane 2: ab314773 IP in Mouse lung tissue lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab314773 in mouse lung tissue lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-RAGE antibody [RM1078] (ab314773) at 1/30 dilution

  All lanes: Mouse lung tissue lysate

  Secondary

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

  Exposure time: 4s

• 

  Immunoprecipitation - Anti-RAGE antibody [RM1078] (ab314773)

  RAGE was immunoprecipitated from 0.35 mg Rat lung tissue lysate with ab314773 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314773 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: Rat lung tissue lysate
  Lane 2: ab314773 IP in Rat lung tissue lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab314773 in rat lung tissue lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-RAGE antibody [RM1078] (ab314773) at 1/30 dilution

  All lanes: Rat lung tissue lysate

  Secondary

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

  Exposure time: 4s