Anti-KIF5B+KIF5C antibody [EPR10276(B)]

10 product images

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  Immunoprecipitation - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  ab167429 (purified) at 1:20 dilution (2μg) immunoprecipitating KIF5B in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.
  
  Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
  Lane 2 (+): ab167429 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab167429 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
  
  For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
  Blocking and diluting buffer: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  Predicted band size: 110 kDa

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  Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  Blocking and diluting buffer: 5% NFDM/TBST

  All lanes: Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429) at 1/5000 dilution

  Lane 1: Jurkat (human acute T cell leukemia) whole cell lysates at 20 µg

  Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell ) whole cell lysates at 20 µg

  Lane 3: Mouse brain lysates at 20 µg

  Lane 4: Rat brain lysates 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: 110 kDa

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  Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  Lane 1: Wild-type HAP1 whole cell lysate (20 μg)
  Lane 2: KIF5B knockout HAP1 whole cell lysate (20 μg)
  

  Lanes 1 - 2: Merged signal (red and green). Green - ab167429 observed at 110 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.

  Unpurified ab167429 was shown to specifically react with KIF5B in wild-type cells as signal was lost in KIF5B knockout cells. Wild-type and KIF5B knockout samples were subjected to SDS-PAGE. ab167429 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  Predicted band size: 110 kDa

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  Flow Cytometry (Intracellular) - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling KIF5B with purified ab167429 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling KIF5B with Purified ab167429 at 1:250 dilution (0.07 μg/ml). Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

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  Immunocytochemistry/ Immunofluorescence - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  Immunocytochemistry/Immunofluorescence analysis HepG2(human hepatocellular carcinoma) labelling KIF5B with purified ab167429 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077). Nuclei counterstained with DAPI (blue).
  

  Control: PBS only

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  Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  All lanes: Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429) at 1/1000 dilution

  Lane 1: Human fetal kidney lysate at 10 µg

  Lane 2: HeLa lysate at 10 µg

  Lane 3: Jurkat lysate at 10 µg

  Lane 4: HepG2 lysate at 10 µg

  Secondary

  All lanes: Goat anti-rabbit HRP at 1/2000 dilution

  Predicted band size: 110 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human brain tissue, labeling KIF5B using unpurified ab167429 at a 1/100 dilution.
  

  Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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  Immunocytochemistry/ Immunofluorescence - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

  Immunofluorescence analysis of HeLa cells, labeling KIF5B using unpurified ab167429 at a 1/100 dilution.

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  Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429)

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

  Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used a GAPDH loading control.

  All lanes: Western blot - Anti-KIF5B+KIF5C antibody [EPR10276(B)] (ab167429) at 1/1000 dilution

  Lane 1: 293T (Human embryonic kidney epithelial cell) +OE-empty at 20 µg

  Lane 2: 293T (Human embryonic kidney epithelial cell) +OE-mouse KIF5A at 20 µg

  Lane 3: 293T (Human embryonic kidney epithelial cell) +OE-mouse KIF5C at 20 µg

  Lane 4: 293T (Human embryonic kidney epithelial cell) +OE-mouse KIF5B 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: 110 kDa

  Exposure time: 1s