Anti-CITED2 antibody [EPR27399-62]

12 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling CITED2 with ab314757 at 1/500 (1.034 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse cerebrum. The section was incubated with ab314757 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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

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  Western blot - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  The expression profile observed is consistent with what has been described in the literature (PMID: 17906695).
  The band beneath the target band (30 kDa) is expected to be degradation products.
  
  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
  Exposure time: Lane 1, 2: 15 seconds Lane 3, 4: 103 seconds

  All lanes: Western blot - Anti-CITED2 antibody [EPR27399-62] (ab314757) at 1/1000 dilution

  Lane 1: Untreated MEF (mouse embryo fibroblast) whole cell lysate at 20 µg

  Lane 2: MEF treated with 5 uM MG-132 for 6 hours, whole cell lysate at 20 µg

  Lane 3: Untreated PC-12 (rat adrenal gland pheochromocytoma ) whole cell lysate at 20 µg

  Lane 4: PC-12 treated with 20 uM MG-132 for 5 hours, 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 1/100000 dilution

  Observed band size: 30 kDa, 36 kDa

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  Dot Blot - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Dot blot analysis of CITED2 using ab314757 at 1:1000 (0.517 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.
  Exposure time: 48 seconds
  Blocking and diluting buffer and concentration: 5% NFDM/TBST

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HT-1080 (human fibrosarcoma epithelial cell) cells labelling CITED2 with ab314757 at 1/50 (10.34 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 nuclear staining in HT-1080 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). 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.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Immunohistochemical analysis of paraffin-embedded mouse pancreatic cancer tissue labeling CITED2 with ab314757 at 1/500 (1.034 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse pancreatic cancer. The section was incubated with ab314757 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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling CITED2 with ab314757 at 1/500 (1.034 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat cerebrum. The section was incubated with ab314757 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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling CITED2 with ab314757 at 1/500 (1.034 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat colon. The section was incubated with ab314757 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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling CITED2 with ab314757 at 1/500 (1.034 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse colon. The section was incubated with ab314757 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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunoprecipitation - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  CITED2 was immunoprecipitated from 0.35 mg MEF (mouse embryo fibroblast) treated with 5 μM MG-132 for 6 hours whole cell lysate with ab314757 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314757 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: MEF (mouse embryo fibroblast) treated with 5 μM MG-132 for 6 hours whole cell lysate
  Lane 2: ab314757 IP in MEF (mouse embryo fibroblast) treated with 5 μM MG-132 for 6 hours whole cell lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab314757 in MEF treated with 5 μM MG-132 for 6 hours whole cell lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-CITED2 antibody [EPR27399-62] (ab314757) at 1/30 dilution

  All lanes: MEF (mouse embryo fibroblast) treated with 5 μM MG-132 for 6 hours whole cell lysate

  Secondary

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

  Exposure time: 3min

• 

  Immunoprecipitation - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  CITED2 was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab314757 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314757 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: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 10 ug
  Lane 2: ab314757 IP in K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab314757 in K-562 whole cell lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-CITED2 antibody [EPR27399-62] (ab314757) at 1/30 dilution

  All lanes: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate

  Secondary

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

  Exposure time: 3min

• 

  Western blot - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  Lysates were freshly made and used immediately to minimize protein degradation.

  All lanes: Western blot - Anti-CITED2 antibody [EPR27399-62] (ab314757) at 1/1000 dilution

  Lane 1: Mouse brain tissue lysate at 20 µg

  Lane 2: Rat brain 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: 30 kDa

  Exposure time: 3min

• 

  Western blot - Anti-CITED2 antibody [EPR27399-62] (ab314757)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  
  Exposure time: Lane 1-3: 48 seconds Lane 4: 37 seconds

  All lanes: Western blot - Anti-CITED2 antibody [EPR27399-62] (ab314757) at 1/1000 dilution

  Lane 1: HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate at 20 µg

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

  Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

  Lane 4: K-562 (human chronic myelogenous leukemia 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 1/100000 dilution

  Observed band size: 30 kDa