Key Product Details

Species Reactivity

Validated:

Mouse, Rat

Cited:

Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all p19ARF/CDKN2A Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide conjugated to KLH, derived from a sequence within residues 50-150 of mouse p19ARF/CDKN2A. [Swiss-Prot #Q64364]

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 31827530).

Localization

Nuclear

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

Novus Biologicals Rabbit p19ARF/CDKN2A Antibody - BSA Free (NB200-106) is a polyclonal antibody validated for use in IHC, WB, ELISA and ICC/IF. Anti-p19ARF/CDKN2A Antibody: Cited in 48 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for p19ARF/CDKN2A Antibody - BSA Free

Western Blot: p19ARF/CDKN2A AntibodyBSA Free [NB200-106]

Western Blot: p19ARF/CDKN2A AntibodyBSA Free [NB200-106]

Western Blot: p19ARF/CDKN2A Antibody [NB200-106] - Analysis of p19ARF/CDKN2A in (A) NIH/3T3 and (B) MEF cells.
Immunocytochemistry/ Immunofluorescence: p19ARF/CDKN2A Antibody - BSA Free [NB200-106]

Immunocytochemistry/ Immunofluorescence: p19ARF/CDKN2A Antibody - BSA Free [NB200-106]

Immunocytochemistry/Immunofluorescence: p19ARF/CDKN2A Antibody [NB200-106] - Antibody was tested in MEF cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).
Immunohistochemistry: p19ARF/CDKN2A Antibody - BSA Free [NB200-106]

Immunohistochemistry: p19ARF/CDKN2A Antibody - BSA Free [NB200-106]

Immunohistochemistry: p19ARF/CDKN2A Antibody [NB200-106] - Adult mouse kidney (glomerulus). Image from verified customer review.
Immunohistochemistry-Paraffin: p19ARF/CDKN2A Antibody - BSA Free [NB200-106]

Immunohistochemistry-Paraffin: p19ARF/CDKN2A Antibody - BSA Free [NB200-106]

Immunohistochemistry-Paraffin: p19ARF/CDKN2A Antibody [NB200-106] - Analysis of FFPE mouse spleen using p19ARF/CDKN2A antibody at 10 ug/mL on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 30 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Whole slide scanning and capturing of representative images was performed using Aperio AT2 (Leica Biosystems). Nuclear staining in the germinal centers of spleen was observed. Staining was performed by Histowiz.

Applications for p19ARF/CDKN2A Antibody - BSA Free

Application
Recommended Usage

ELISA

1:100-1:2000

Immunocytochemistry/ Immunofluorescence

1:50 - 1:100

Immunohistochemistry

10 ug/ml

Immunohistochemistry-Frozen

1 ug/ml

Immunohistochemistry-Paraffin

10 ug/ml

Western Blot

2 ug/ml
Application Notes
In WB a band can be seen at approximately 19 kDa. 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.

Reviewed Applications

Read 1 review rated 3 using NB200-106 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% 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

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: p19ARF/CDKN2A

p19ARF (mouse) and p14ARF (human) has become an important player in cell cycle regulation. In mice, tumor suppressor effects appear to be mediated by interactions between p19ARF and the p53 tumor suppressor protein. p19ARF counters uncontrolled proliferation and oncogenic signals in p53 dependent pathways. Proteins encoded by the INK4a/ARF locus also play a role in Abelson virus transformation. Both p16INK4a and p19ARF are expressed in many cells as they emerge from the apoptotic crisis that characterizes the transformation process. p19ARF is an important part in cellular defense against transforming signals from the Abl oncoprotein, providing direct evidence that the p19ARF-p53 regulatory loop plays an important role in lymphoma induction.

Alternate Names

alternative reading frame, ARF, ARF-INK4a, CDK4I, CDKN2A, Cyclin-dependent kinase 4 inhibitor A, cyclin-dependent kinase inhibitor 2A, cyclin-dependent kinase inhibitor 2A (p16, inhibits CDK4), cyclin-dependent kinase inhibitor 2A, isoform 3, cyclin-dependent kinase inhibitor 2A, isoforms 1/2, cyclin-dependent kinase inhibitor protein, INK4, INK4a, Ink4a/Arf, INK4a-ARF, mitochondrial smARF', MTS1, p14, p14ARF, P16, p16(INK4a), p16-INK4, P16INK4A, p16-INK4a, p19, p19, p19ARF, Pctr1

Gene Symbol

Cdkn2a

Additional p19ARF/CDKN2A Products

Product Documents for p19ARF/CDKN2A Antibody - BSA Free

Certificate of Analysis

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

Download SDS

Product Specific Notices for p19ARF/CDKN2A 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 p19ARF/CDKN2A Antibody - BSA Free

Customer Reviews for p19ARF/CDKN2A Antibody - BSA Free (1)

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Customer Images

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  • p19ARF Antibody
    Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: Adult kidney
    Species: Mouse
    Verified Customer | Posted 05/07/2018
    mouse glomerulus
    p19ARF/CDKN2A Antibody - BSA Free NB200-106

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Protocols

View specific protocols for p19ARF/CDKN2A Antibody - BSA Free (NB200-106):



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 0.5% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS with 0.1% Tween 20. 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.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow proper laboratory procedures for the disposal of formalin.

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

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FAQs for p19ARF/CDKN2A Antibody - BSA Free

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  • Q: Is there a recommended antigen retrieval method for this product for use in IHC-P?

    A: For NB200-106 I would recommend our standard HIER Citrate Buffer pH 6.0.

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