OCT4 Antibody - BSA Free

Novus Biologicals | Catalog # NB100-2379

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Bovine, Primate

Cited:

Human, Mouse, Rat, Bovine

Predicted:

Feline (93%), Porcine (93%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Knockout Validated, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC, Knockdown Validated

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all OCT4 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal portion of the human OCT4 protein (between residues 100-200) [UniProt# Q01860]

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: Feline (93%), Porcine (93%). Customer feedback suggests that this antibody does not give good results in IHC-P on feline tissues. Bovine reactivity reported in scientific literature (PMID: 23054358).

Localization

Nuclear

Marker

Embryonic Stem Cell Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

40 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for OCT4 Antibody - BSA Free

Western Blot: OCT4 AntibodyBSA Free [NB100-2379]

Western Blot: OCT4 AntibodyBSA Free [NB100-2379]

OCT4-Antibody-Western-Blot-NB100-2379-img0014.jpg
Western Blot: OCT4 AntibodyBSA Free [NB100-2379]

Western Blot: OCT4 AntibodyBSA Free [NB100-2379]

Western Blot: OCT4 Antibody [NB100-2379] - WB detection of OCT4 in mouse brain lysate using OCT4 antibody with ECL based detection. The antibody generated a specific band of OCT4 at the expected ~40kDa position.
Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody [NB100-2379] - An intracellular stain was performed on Jurkat with NB100-2379 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody.
Immunocytochemistry/ Immunofluorescence: OCT4 Antibody - BSA Free [NB100-2379]

Immunocytochemistry/ Immunofluorescence: OCT4 Antibody - BSA Free [NB100-2379]

Immunocytochemistry/Immunofluorescence: OCT4 Antibody [NB100-2379] - Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- NB100-2379 at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Western Blot: OCT4 AntibodyBSA Free [NB100-2379]

Western Blot: OCT4 AntibodyBSA Free [NB100-2379]

Western Blot: OCT4 Antibody [NB100-2379] - MDA-MB-231 subclones transfected with NTC or HIF-1 alpha shRNA vector were treated with vehicle (V) or paclitaxel (P) and Western blot assay was performed. WB image submitted by a verified customer review.
Immunohistochemistry-Paraffin: OCT4 Antibody - BSA Free [NB100-2379]

Immunohistochemistry-Paraffin: OCT4 Antibody - BSA Free [NB100-2379]

Immunohistochemistry-Paraffin: OCT4 Antibody [NB100-2379] - Analysis of a FFPE tissue section of mouse brain using 1:200 dilution of OCT4 antibody. The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.
Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379] - An intracellular stain was performed on U2-OS cells with OCT4 NB100-2379AF647 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.
Western Blot: OCT4 AntibodyBSA Free [NB100-2379]

Western Blot: OCT4 AntibodyBSA Free [NB100-2379]

Western Blot: OCT4 Antibody [NB100-2379] - The expression of pluripotency factors Oct4 in human head and neck cancer cell lines.
Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody [NB100-2379] - An intracellular stain was performed on Jurkat cells with OCT4 Antibody NB100-2379 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).
Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody [NB100-2379] - An intracellular stain was performed on Neuro2a cells with OCT4 Antibody NB100-2379 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).
Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody - BSA Free [NB100-2379]

Flow Cytometry: OCT4 Antibody [NB100-2379] - An intracellular stain was performed on Ntera2 cells with OCT4 Antibody NB100-2379 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).
OCT4 Antibody - BSA Free

OCT4 in U-2 OS Human Cell Line.

OCT4 was detected in immersion fixed U-2 OS human osteosarcoma cell line using Rabbit anti-OCT4 Affinity Purified Polyclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB100-2379AF488) (green) at 10 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
OCT4 Antibody - BSA Free

Detection of OCT4 in A431 Human Cell Line by Flow Cytometry.

An intracellular stain was performed on A431 human skin carcinoma cell line using Rabbit anti-OCT4 Affinity Purified Polyclonal Antibody conjugated to DyLight 650 (Catalog # NB100-2379C, blue histogram) or matched control antibody (orange histogram) at 2.5 µg/mL for 30 minutes at RT.
OCT4 Antibody - BSA Free

Western Blot: OCT4 Antibody - BSA Free [NB100-2379] -

Western Blot: OCT4 Antibody - BSA Free [NB100-2379] - Protein expression of NANOG a, OCT4 b & SOX2 c in bovine uterine tissues obtained from control cows & from cows with adenomyosis. Data were normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Bars represent the mean ± SEM. There were no statistical differences between uterine normal & adenomyotic tissues (P > 0.05), as determined by Student’s t-test. Representative blots for NANOG, OCT4, SOX2 & GAPDH are shown below the graphs d. MM – molecular weight marker, C – tissues obtained from control cows, ADENO – tissues obtained from cows with adenomyosis Image collected & cropped by CiteAb from the following publication (http://www.rbej.com/content/13/1/110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
OCT4 Antibody - BSA Free

Western Blot: OCT4 Antibody - BSA Free [NB100-2379] -

Western Blot: OCT4 Antibody - BSA Free [NB100-2379] - Protein expression of pluripotency markers in uterine cells isolated from control cows & from cows with adenomyosis. NANOG a OCT4 b & SOX2 c protein expression in uterine stromal cells. NANOG d OCT4 e & SOX2 f protein expression in uterine myometrial cells. Data were normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Bars represent the mean ± SEM. Asterisks indicate statistical differences between uterine normal & adenomyotic tissue (*P < 0.05; **P < 0.01), as determined by Student’s t-test. Representative blots for NANOG, OCT4, SOX2 & GAPDH are shown below the graphs g. MM – molecular weight marker, C – cells obtained from control cows, ADENO – cells obtained from cows with adenomyosis Image collected & cropped by CiteAb from the following publication (http://www.rbej.com/content/13/1/110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
OCT4 Antibody - BSA Free

Detection of OCT4 in U-2 OS Human Cell Line by Flow Cytometry.

An intracellular stain was performed on U-2 OS human osteosarcoma cell line with Rabbit anti-OCT4 Affinity-purified Polyclonal Antibody conjugated to Phycoerythrin (Catalog # NB100-2379PE, blue histogram) or matched control antibody (Catalog # NBP2-24983, orange histogram) at 2.5 µg/mL for 30 minutes at RT.

Applications for OCT4 Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

1:50-1:200

Immunocytochemistry/ Immunofluorescence

1:50-1:200

Immunohistochemistry

1:200. Use reported in scientific literature (PMID 30726734)

Immunohistochemistry-Paraffin

1:200

Western Blot

reported by customer review
Application Notes
In Western blot, a band is seen at ~40 kDa.

Reviewed Applications

Read 2 reviews rated 5 using NB100-2379 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: OCT4

OCT4 (octamer-binding protein 4) is a transcription factor that binds to the octamer motif (5'-ATTTGCAT-3') and generates a trimeric complex with SOX2 on DNA for controlling the expression of several genes (YES1, FGF4, UTF1, ZFP206 etc.) involved in embryonic development. Expressed mainly in the nucleus, OCT4 interacts with UBE2I, ZSCAN10, PKM, WWP2 etc. and OCT4-PKM interaction positively regulate OCT4's transcriptional activity. Once translated, its modifications such as sumoylation enhances the protein stability, DNA binding and transactivation activity and is required for enhanced YES1 expression. OCT4 undergoes 'Lys-63'-linked polyubiquitination by WWP2 leading to proteasomal degradation. OCT4 is found in oocytes and is zygotically expressed in pluripotent cells of pre-gastrulation embryos, and has been observed to be essential for early embryogenesis as well as for embryonic stem cell pluripotency. OCT4 is expressed in developing brain with highest levels in specific cell layers of the cortex, the olfactory bulb, hippocampus and cerebellum. Its expression decreases gradually after embryoid body formation, however, improper re-activation of OCT4 contributes to oncogenic processes and it has been implicated in certain malignancies such as breast and ovarian cancer.

Alternate Names

class 5, transcription factor 1, MGC22487, Oct-3, OCT3Oct4, Oct-4, Octamer-binding protein 3, otc-4, OTF3POU domain class 5, transcription factor 1, OTF4, POU class 5 homeobox 1, POU-type homeodomain-containing DNA-binding protein

Entrez Gene IDs

5460 (Human); 18999 (Mouse)

Gene Symbol

POU5F1

UniProt

Q01860

Additional OCT4 Products

Product Documents for OCT4 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for OCT4 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for OCT4 Antibody - BSA Free

Customer Reviews for OCT4 Antibody - BSA Free (2)

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Customer Images

Showing  1 - 2 of 2 reviews Showing All
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  • OCT4 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Breast cancer cells
    Species: Human
    Verified Customer | Posted 05/18/2020
    MDA-MB-231 subclones transfected with NTC or HIF-1 alpha shRNA vector were treated with vehicle (V) or paclitaxel (P) and immunoblot assay was performed.
    OCT4 Antibody - BSA Free NB100-2379
  • OCT4 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: human head and neck cancer cell lines
    Species: Human
    Verified Customer | Posted 10/11/2018
    The expression of pluripotency factors Oct4 in human head and neck cancer cell lines.
    OCT4 Antibody - BSA Free NB100-2379

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Protocols

View specific protocols for OCT4 Antibody - BSA Free (NB100-2379):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer all the time).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for OCT4 Antibody - BSA Free

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  • Q: We are looking for the following markers to use to stain cells derived from both human and mouse embryonic and induced pluripotent stem cells: undifferentiated cells, neurons, oligodendrocytes, and astrocytes.

    A: For ES markers some popular targets are OCT4 (NB100-2379), KLF4 (NBP2-17070) and SOX2 (NB110-37235). PEA15 is expressed in Astrocytes and this might be one of interest for you (NBP1-61390). OCT4 is one that can also be expressed in Oligodendrocytes.

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