Anti-RFP antibody [EPR28246-45] - BSA and Azide free

8 product images

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  Immunoprecipitation - Anti-RFP antibody [EPR28246-45] - BSA and Azide free (ab314226)

  This data was developed using Anti-RFP antibody [EPR28246-45] ab314225, the same antibody clone in a different buffer formulation.
  RFP was immunoprecipitated from 0.35 mg 293T cells transfected with a red fluorescent protein expression vector containing a His-tag, whole cell lysate with Anti-RFP antibody [EPR28246-45] ab314225 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-RFP antibody [EPR28246-45] ab314225 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: 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag, whole cell lysate
  Lane 2: Anti-RFP antibody [EPR28246-45] ab314225 IP in 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag, whole cell lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-RFP antibody [EPR28246-45] ab314225 in 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag, whole cell lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-RFP antibody [EPR28246-45] (Anti-RFP antibody [EPR28246-45] ab314225) at 1/30 dilution

  All lanes: 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag, whole cell lysate

  Secondary

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

  RFP was immunoprecipitated from 0.35 mg 293T cells transfected with a red fluorescent protein expression vector containing a His-tag, whole cell lysate with Anti-RFP antibody [EPR28246-45] ab314225 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-RFP antibody [EPR28246-45] ab314225 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: 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag, whole cell lysate
  Lane 2: Anti-RFP antibody [EPR28246-45] ab314225 IP in 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag, whole cell lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-RFP antibody [EPR28246-45] ab314225 in 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag, whole cell lysate
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

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  Flow Cytometry (Intracellular) - Anti-RFP antibody [EPR28246-45] - BSA and Azide free (ab314226)

  This data was developed using Anti-RFP antibody [EPR28246-45] ab314225, the same antibody clone in a different buffer formulation.
  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells transfected with a red fluorescent protein expression vector containing a His-tag (upper right) / 293T cells transfected with an empty vector containing a His tag (low right) cells labelling RFP with Anti-RFP antibody [EPR28246-45] ab314225 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Immunocytochemistry/ Immunofluorescence - Anti-RFP antibody [EPR28246-45] - BSA and Azide free (ab314226)

  This data was developed using Anti-RFP antibody [EPR28246-45] ab314225, the same antibody clone in a different buffer formulation.
  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling RFP with Anti-RFP antibody [EPR28246-45] ab314225 at 1/2000 (0.252 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) antibody at 1/500 (4 ug/ml) dilution (Green). Confocal image showing positive staining in 293T cells transfected with a RFP expression vector containing a Myc-tag. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Myc-Tag Mouse mAb (Alexa Fluor® 488) was used to counterstain at 1/100 (0.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) at 1/500 (4 ug/ml) dilution.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RFP antibody [EPR28246-45] - BSA and Azide free (ab314226)

  This data was developed using Anti-RFP antibody [EPR28246-45] ab314225, the same antibody clone in a different buffer formulation.
  Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling RFP with Anti-RFP antibody [EPR28246-45] ab314225 at 1/2000 (0.252 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on rat kidney. The section was incubated with Anti-RFP antibody [EPR28246-45] ab314225 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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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RFP antibody [EPR28246-45] - BSA and Azide free (ab314226)

  This data was developed using Anti-RFP antibody [EPR28246-45] ab314225, the same antibody clone in a different buffer formulation.
  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling RFP with Anti-RFP antibody [EPR28246-45] ab314225 at 1/2000 (0.252 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on mouse kidney. The section was incubated with Anti-RFP antibody [EPR28246-45] ab314225 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-RFP antibody [EPR28246-45] - BSA and Azide free (ab314226)

  This data was developed using Anti-RFP antibody [EPR28246-45] ab314225, the same antibody clone in a different buffer formulation.
  Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling RFP with Anti-RFP antibody [EPR28246-45] ab314225 at 1/2000 (0.252 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on human kidney. The section was incubated with Anti-RFP antibody [EPR28246-45] ab314225 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-RFP antibody [EPR28246-45] - BSA and Azide free (ab314226)

  This data was developed using Anti-RFP antibody [EPR28246-45] ab314225, the same antibody clone in a different buffer formulation.
  Immunohistochemical analysis of paraffin-embedded Panel A HEK-293T cells transfected with a Red fluorescent protein expression vector containing a his tag. Panel B HEK-293T cells transfected with empty vector containing a his tag. Tissue labeling RFP with Anti-RFP antibody [EPR28246-45] ab314225 at 1/2000 (0.252 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) HEK-293T transfected with a red fluorescent protein expression vector and no staining on (B) HEK-293T transfected with empty vector. The section was incubated with Anti-RFP antibody [EPR28246-45] ab314225 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-RFP antibody [EPR28246-45] - BSA and Azide free (ab314226)

  This data was developed using Anti-RFP antibody [EPR28246-45] ab314225, the same antibody clone in a different buffer formulation.
  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
  In Western blot, Anti-6X His tag antibody [EPR20547]-CHIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-RFP antibody [EPR28246-45] (Anti-RFP antibody [EPR28246-45] ab314225) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a His tag, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag, 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 1/20000 dilution

  Observed band size: 35 kDa

  Exposure time: 15s

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
  In Western blot, Anti-6X His tag antibody [EPR20547]-CHIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.