Anti-Oct4 antibody [EPR17929] - Low endotoxin, Azide free

8 product images

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  Immunoprecipitation - Anti-Oct4 antibody [EPR17929] - Low endotoxin, Azide free (ab215386)

  Oct4 was immunoprecipitated from 1mg of NCCIT (Human pluripotent embryonic carcinoma) whole cell extract with Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 at 1/50 dilution. Western blot was performed from the immunoprecipitate using Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 at 1/10000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

  Lane 1: NCCIT whole cell extract 10 µg (Input). Lane 2: Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 IP in NCCIT whole cell extract. Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 in NCCIT whole cell extract.
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  Exposure time: 3 minutes

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557).

  All lanes: Immunoprecipitation - Anti-Oct4 antibody [EPR17929] - ChIP Grade (Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557)

  Predicted band size: 38 kDa

  Observed band size: 45 kDa

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  Flow Cytometry (Intracellular) - Anti-Oct4 antibody [EPR17929] - Low endotoxin, Azide free (ab215386)

  Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 staining OCT-4 in the human cell line NCCIT (human pluripotent embryonal carcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/70. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody (Red).
  

  Isotype control: Rabbit IgG monoclonal [EPR25A] Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 (Black).

  Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557).

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  Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17929] - Low endotoxin, Azide free (ab215386)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (Human pluripotent embryonic carcinoma) cells (positive cell line) or NIH/3T3 (Mouse embyro fibroblast) cells (negative cell line) labeling Oct4 with Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on NCCIT cell line. Negative expression in NIH/3T3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
  The negative controls are as follows:
  -ve control 1: Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
  -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557).

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  This image is courtesy of a customer review submitted by Joanne Lacey.

  Immunocytochemistry/ Immunofluorescence - Anti-Oct4 antibody [EPR17929] - Low endotoxin, Azide free (ab215386)

  Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 staining Oct4 in Human embryonic stem cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with formaldehyde , permeabilized with 0.1% Triton in PBS for 1 hour and blocked with 10% Serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/200 in PBS with 0.1% Tween20) for 16 hours at 4°C. A monoclonal Goat Anti-rabbit Alexa Fluor^® 594  was used as the secondary antibody at 1/200 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17929] - Low endotoxin, Azide free (ab215386)

  Immunohistochemical analysis of paraffin-embedded Human dysgerminoma of ovary tissue labeling Oct4 with Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution. Nuclear and weak cytoplasmic staining on cancer cells of Human dysgerminoma of ovary is observed. Counter stained with Hematoxylin.
  

  Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557).

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17929] - Low endotoxin, Azide free (ab215386)

  Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Oct4 with Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution. Negative staining on Human breast cancer. Counter stained with Hematoxylin.
  

  Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557).

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Oct4 antibody [EPR17929] - Low endotoxin, Azide free (ab215386)

  Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Oct4 with Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution. Negative staining on adult Human testis. Counter stained with Hematoxylin.
  

  Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557).

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

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  ChIP-sequencing - Anti-Oct4 antibody [EPR17929] - Low endotoxin, Azide free (ab215386)

  Chromatin was prepared from NCCIT (Human pluripotent embryonic carcinoma cell line) cells. ChIP was performed with 10^7 NCCIT cells and 8 µg of Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557 [EPR17929]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Oct4 antibody [EPR17929] - ChIP Grade ab181557).