CENPF Antibody - BSA Free

Novus Biologicals | Catalog # NB500-101

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Bovine, Equine

Cited:

Human, Mouse, Bovine, Equine

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunohistochemistry Whole-Mount, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Knockdown Validated

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunohistochemistry Whole-Mount, Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all CENPF Primary Antibodies »

Product Specifications

Immunogen

A bacterial fusion protein from the C-terminus of human CENPF. [UniProt# P49454]

Localization

Nucleus

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for CENPF Antibody - BSA Free

Knockdown Validated: CENPF Antibody - BSA Free [NB500-101]

Immunocytochemistry/ Immunofluorescence: CENPF Antibody - BSA Free [NB500-101]

CENPF-Antibody-Immunohistochemistry-Paraffin-NB500-101-img0009.jpg
Western Blot: CENPF AntibodyBSA Free [NB500-101]

Western Blot: CENPF AntibodyBSA Free [NB500-101]

Western Blot: CENPF Antibody [NB500-101] - 25ug of asynchronous and mitotically blocked HeLa lysates were loaded in each respective lane.
Immunocytochemistry/ Immunofluorescence: CENPF Antibody - BSA Free [NB500-101]

Immunocytochemistry/ Immunofluorescence: CENPF Antibody - BSA Free [NB500-101]

Immunocytochemistry/Immunofluorescence: CENPF Antibody [NB500-101] - Distribution of CENP-F (red), largely associated with the mitotic spindle and a clearly distinguishable frac.
Immunocytochemistry/ Immunofluorescence: CENPF Antibody - BSA Free [NB500-101]

Immunocytochemistry/ Immunofluorescence: CENPF Antibody - BSA Free [NB500-101]

Immunocytochemistry/Immunofluorescence: CENPF Antibody [NB500-101] - CENPF antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
Immunocytochemistry/ Immunofluorescence: CENPF Antibody - BSA Free [NB500-101]

Immunocytochemistry/ Immunofluorescence: CENPF Antibody - BSA Free [NB500-101]

Immunocytochemistry/Immunofluorescence: CENPF Antibody [NB500-101] - Confocal immunofluorescent analysis of HeLa cells using CENPF antibody (NB500-101, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green, A). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red, B). DAPI was used to stain the cell nuclei (blue, C).
CENPF Antibody - BSA Free

Western Blot: CENPF Antibody - BSA Free [NB500-101] -

Requirements for CENP-F kinetochore localization.A, representative images of mitotic HeLa cells electroporated with mCherry, mCherry-CENP-F2688-C WT, or mCherry-CENP-F2688-C/C3207A (farnesylation mutant). Scale bar, 5 μm. As for CENP-E, both the WT and the unfarnesylated mutant CENP-F constructs localize at kinetochore. B, mCherry-CENP-F2688-C sample was visualized by EM after glycerol spraying and low-angle platinum shadowing (right panel). The elongated shape of the observed particles is consistent with the secondary structure expected for the mCherry-tag coiled-coil construct (right panel). C and D, SEC elution profiles and SDS-PAGE analysis of binding experiments with 16 μm each of mCherry-CENP-F2688-C WT (C) or C2961S mutant (D) and 4 μm BUB1FL/BUB3 complex. The shift in elution volume of BUB1/BUB3 is observed with both WT and mutant CENP-F, but it is significantly less pronounced for the CENP-F mutant, suggesting that the C2961S mutation reduces the affinity of CENP-F for the BUB1 kinase domain without completely abolishing it. E, SEC elution profile and SDS-PAGE analysis of a binding experiment with 16 μm each of mCherry-CENP-F2688-C and eGFP-CENP-E2070-C. No shift is observed, indicating that the tested constructs do not interact. The elution profile and SDS-PAGE of eGFP-CENP-E2070-C WT in E is the same already shown in Fig. 3, B and D. Similarly, the elution profiles and SDS-PAGE of mCherry-CENP-F2688-C in E is the same as in Fig. 4, D, E, and G. Similarly, the elution profiles and SDS-PAGE of BUB1/BUB3 in C and D are the same. These repetitions were included to facilitate the interpretation of binding experiments by inclusion of elution references. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29748388), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
CENPF Antibody - BSA Free

Immunohistochemistry: CENPF Antibody - BSA Free [NB500-101] -

Kinetochore localization of RZZ and MAD1 are independent of CENP-E and CENP-F.A–H, representative images and quantification of protein kinetochore levels in HeLa cells mock-treated or depleted of Zwilch (A), CENP-E (B, E, and H), CENP-F (C, F, and G), or co-depleted of CENP-E and CENP-F (D). Scale bar, 10 μm. Zwilch depletion does not affect the localization of CENP-E (A). CENP-E depletion does not affect the localization of Zwilch (B), MAD1 (E), and CENP-F (H). Similarly, CENP-F depletion does not interfere with the recruitment of Zwilch (C), MAD1 (F), and CENP-E (G). Co-depletion of CENP-E and CENP-F has no effects on localization of Zwilch (D). The graphs show mean intensity of one (B, C, and E), two (D and F), or three (A, G, and H) experiments; the error bars indicate S.E., and the mean values for nondepleted cells are set to 1. Elements in the left column of A (negative controls of the RNAi experiments) are also shown in Fig. S3C. Elements in G are shown again in Fig. S2E. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29748388), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for CENPF Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

1:50-1:200

Immunocytochemistry/ Immunofluorescence

1:200-1:1000immunoprecipitation 1:50

Immunohistochemistry Whole-Mount

reported in scientific literature (PMID 24876181)

Immunohistochemistry-Frozen

reported in scientific literature (PMID 26195156)

Immunohistochemistry-Paraffin

reported in scientific literature (PMID 20398247)

Immunoprecipitation

1:50

Western Blot

1:1500-1:2500
Application Notes
In Western Blot, a band can be seen at ~330 kDa. In ICC/IF, nuclear staining was observed in HeLa cells.

Reviewed Applications

Read 2 reviews rated 5 using NB500-101 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: CENPF

CENPF interacts with the centromere-kinetochore complex. CENP-F also shows a cell-cycle distribution that is both temporally regulated and diverse in terms of the structural components it localizes to and therefore may play a role in several mitotic events.

Alternate Names

AH antigen, cell-cycle-dependent 350K nuclear protein, CENF, CENP-F, CENP-F kinetochore protein, centromere protein F, centromere protein F (350/400kD, mitosin), centromere protein F, 350/400ka (mitosin), centromere protein F, 350/400kDa (mitosin), hcp-1, Kinetochore protein CENPF, mitosin, PRO1779

Gene Symbol

CENPF

Additional CENPF Products

Product Documents for CENPF 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 CENPF 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 CENPF Antibody - BSA Free

Customer Reviews for CENPF Antibody - BSA Free (2)

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Customer Images

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  • Name: Patrizia Lavia
    Application: Immunofluorescence
    Sample Tested:
    Species: Human
    Verified Customer | Posted 03/24/2015
    The image shows the distribution of CENP-F (red), largely associated with the mitotic spindle and a clearly distinguishable frac
    CENPF Antibody - BSA Free NB500-101
  • Name: Bryan Tinsley
    Application: Immunocytochemistry
    Sample Tested:
    Species: Human
    Verified Customer | Posted 06/05/2014
    Confocal immunofluorescent analysis of HeLa cells using CENPF antibody (NB500-101, 1:5).
    CENPF Antibody - BSA Free NB500-101

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Protocols

View specific protocols for CENPF Antibody - BSA Free (NB500-101):


Western Blot Procedure

1) Resolve aliquots (30 mg) of total HeLa cell lysate on a 8.5% lo-crosslinker gel (110:1, acryl:bis) for efficient separation and transfer.
2) Transfer to nitrocellulose membranes in 20 mM Tris-HCL (pH 8.0)/150 mM glycine/20% (vol/vol) methanol. Transfer time for a 1 mm thick gel is 1 hour 15 minutes at 450mA.
3) Block membranes for 1 hour with 5% (vol/vol) nonfat dry milk/TBS-T (20 mM Tris-HCL, pH 7.6/ 137 mM NaCl/0.1% TWEEN 20)
4) Incubate membranes for 1 hour at room temperature (RT) in NB 500-101 diluted 1:3000 in nonfat dry milk/TBS-T.
5) Wash 3X30 minutes at RT with TBS-T.
6) Incubate membranes with alkaline phosphatase conjugated anti-rabbit IgG for 1 hour (RT).
7) Wash 3 x 30 minutes at RT with TBS-T
8) Develop with ECL reagents (Amersham) and autoradiography.


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FAQs for CENPF Antibody - BSA Free

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  • Q: Could you please check whether you can provide which region/sequence this antibody is raised against in the C-terminus of CENPF?

    A: Unfortunately, I am not able to provide sequence information further than what we have published. I apologize for the inconvenience.

  • Q: What residues of human CENP-F was NB500-101 generated against?

    A: The exact immunogen sequence is proprietary and cannot be released.

  • Q: Could you please check whether you can provide which region/sequence this antibody is raised against in the C-terminus of CENPF?

    A: Unfortunately, I am not able to provide sequence information further than what we have published. I apologize for the inconvenience.

  • Q: What residues of human CENP-F was NB500-101 generated against?

    A: The exact immunogen sequence is proprietary and cannot be released.

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