Anti-IL-13 receptor alpha 2 antibody [EPR22978-163]

6 product images

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  Western blot - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044)

  Lanes 1 - 4: Merged signal (red and green). Green - ab260044 observed at 49 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
  

  ab260044 was shown to react with IL-13 receptor alpha 2 in wild-type A375 cells in western blot with loss of signal observed in IL13RA2 knockout cell line ab273371 (knockout cell lysate Human IL13RA2 knockout A375 cell lysate ab275532). Wild-type and IL13RA2 knockout A375 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab260044 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044) at 1/1000 dilution

  Lane 1: Wild-type A375 cell lysate at 30 µg

  Lane 2: IL13RA2 knockout A375 cell lysate at 30 µg

  Lane 2: Western blot - Human IL13RA2 knockout A375 cell line (Human IL13RA2 knockout A375 cell line ab273381)

  Lane 3: U-87-MG cell lysate at 30 µg

  Lane 4: Daudi cell lysate at 30 µg

  Performed under reducing conditions.

  Predicted band size: 44 kDa

  Observed band size: 49 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044)

  ab260044 staining IL-13 receptor alpha 2 in wild-type A375 cells (top panel) and IL13RA2 knockout A375 cells (bottom panel) (Human IL13RA2 knockout A375 cell line ab273381). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab260044 at 1/500 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

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  Immunoprecipitation - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044)

  IL-13 receptor alpha 2 was immunoprecipitated from 0.35 mg A375 (Human malignant melanoma epithelial cell) whole cell lysate 10ug with ab260044 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab260044 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution. Lane 1: A375 (Human malignant melanoma epithelial cell) whole cell lysate 10ug

  Lane 2: ab260044 IP in A375 whole cell lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab260044 in A375 whole cell lysate

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

  Exposure time: 30 seconds

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab260044).

  All lanes: Immunoprecipitation - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044)

  Predicted band size: 44 kDa

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  Western blot - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044)

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

  The molecular weight observed is consistent with what has been described in the literature (PMID: 30718742).

  Negative control: Daudi (PMID: 30718742).

  Exposure time: Lane 1: 3 minutes; Lane 2: 10 seconds; Lanes 3-4: 3 minutes.

  All lanes: Western blot - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044) at 1/1000 dilution

  Lane 1: Human testis lysate at 20 µg

  Lane 2: A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: A-172 (human brain glioblastoma) whole cell lysate at 20 µg

  Lane 4: Daudi (human burkitt's lymphoma lymphoblast) whole cell 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 1/25000 dilution

  Predicted band size: 44 kDa

  Observed band size: 55 kDa, 65 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A375 (human malignant melanoma epithelial cell) cells labelling IL-13 receptor alpha 2 with ab260044 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). UNABLE TO ENCODE THE VARIABLE PLEASE DO MANUALLY is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab260044).

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  Indirect ELISA - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (ab260044)

  ELISA analysis of IL13RA2 recombinant protein at 1000 ng/mL with ab260044. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.