MDM2/HDM2 Antibody (SMP14) - BSA Free

Novus Biologicals | Catalog # NB100-2736

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunoprecipitation

Cited:

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

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # SMP14

Format

BSA Free
View all MDM2/HDM2 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide C-SRPSTSSRRRAISE, corresponding to amino acids 154-167 of human MDM2/HDM2. [UniProt# Q00987]

Epitope

Amino acids 154 - 167.

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 15625077). Mouse reactivity reported in scientific literature (PMID: 25981963).

Localization

Nuclear and cytoplasmic (expressed predominantly in the nucleoplasm).

Specificity

The SMP14 monoclonal antibody also recognises a peptide epitope around Thr-216 of murine MDM2 (when Thr-216 is unphosphorylated) see Zhang & Prives, 2001 (PMID: 11359766) for further details. SMP14 also cross reacts with some cytokeratins (6, 14 & 16). This is only a problem when working with certain epithelial cells and not fibroblasts.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for MDM2/HDM2 Antibody (SMP14) - BSA Free

Western Blot: MDM2/HDM2 Antibody (SMP14)BSA Free [NB100-2736]

Western Blot: MDM2/HDM2 Antibody (SMP14)BSA Free [NB100-2736]

Western Blot: MDM2/HDM2 Antibody (SMP14) [NB100-2736] - Daudi whole cell protein was separated by SDS-PAGE on a 7.5% gel and transferred to PVDF membrane. The membrane was probed with anti-MDM2 antibody at 2 ug/ml and detected with an anti-mouse HRP secondary antibody using chemiluminescence.
Immunohistochemistry-Paraffin: MDM2/HDM2 Antibody (SMP14) - BSA Free [NB100-2736]

Immunohistochemistry-Paraffin: MDM2/HDM2 Antibody (SMP14) - BSA Free [NB100-2736]

Immunohistochemistry-Paraffin: MDM2/HDM2 Antibody (SMP14) [NB100-2736] - Analysis of tissue section of human breast cancer xenograft using MDM2/HDM2 antibody (clone SMP14) at 1:100 dilution. Several of the cancer cells developed a strong nuclear with weak cytoplasmic immunostaining of MDM2.
Immunohistochemistry-Paraffin: MDM2/HDM2 Antibody (SMP14) - BSA Free [NB100-2736]

Immunohistochemistry-Paraffin: MDM2/HDM2 Antibody (SMP14) - BSA Free [NB100-2736]

Immunohistochemistry-Paraffin: MDM2/HDM2 Antibody (SMP14) [NB100-2736] - Analysis of tissue section of human breast cancer xenograft using MDM2/HDM2 antibody (clone SMP14) at 1:100 dilution. Several of the cancer cells developed a strong nuclear with weak cytoplasmic immunostaining of MDM2.

Applications for MDM2/HDM2 Antibody (SMP14) - BSA Free

Application
Recommended Usage

Immunohistochemistry

1:100-1:500

Immunohistochemistry-Frozen

1:200-1:500

Immunohistochemistry-Paraffin

1:100-1:500

Immunoprecipitation

1:10-1:500

Western Blot

1:500
Application Notes
This antibody has also been reported useful in IHC on resin tissue sections. Staining of IHC-P tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 min.

Reviewed Applications

Read 1 review rated 4 using NB100-2736 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 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: MDM2/HDM2

MDM2, also known as HDM2, is a protein of the E3 ubiquitin-ligase family. MDM2 is found predominantly in the nucleus, most localized to the nucleoplasm. MDM2 is over expressed in a wide range of human malignancies including soft tissue carcinomas and breast cancer. MDM2 ubiquinates or contributes to the ubiquination of a variety of proteins, including p53, thereby contributing to their proteosomal degradation. MDM2's interaction with p53 is inhibitory, causing an arrest of the cell cycle and deregulation of apoptosis, which explains MDM2's presence in many cancers. MDM2 is induced by damage to the DNA, and as a result has many isoforms, most of which are present exclusively in cancerous tissues, and non-existent in normal tissues.

Long Name

Double Minute 2 Protein

Alternate Names

HDM2

Entrez Gene IDs

4193 (Human)

Gene Symbol

MDM2

UniProt

Q00987

Additional MDM2/HDM2 Products

Product Documents for MDM2/HDM2 Antibody (SMP14) - 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 MDM2/HDM2 Antibody (SMP14) - 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 MDM2/HDM2 Antibody (SMP14) - BSA Free

Customer Reviews for MDM2/HDM2 Antibody (SMP14) - BSA Free (1)

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Hela whole cell lysate
    Species: Human
    Verified Customer | Posted 09/08/2010

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Protocols

View specific protocols for MDM2/HDM2 Antibody (SMP14) - BSA Free (NB100-2736):

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 at all times).

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 manufacturer's instructions.

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

FAQs for MDM2/HDM2 Antibody (SMP14) - BSA Free

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  • Q: I would like to know the epitope of the antibody HDM2 antibody SMP14 for human.

    A: We have not mapped the exact epitope for NB100-2736, but the immunogen is listed as a the synthetic peptide C-SRPSTSSRRRAISE, corresponding to amino acids 154-167 of human MDM2/HDM2. [UniProt# Q00987]

  • Q: I would like to know where I can expect to see the signals with NB100-2736 in WB. After checking the information from UniProt, the MW of MDM2 seems to be 55 KDa. However, it may be changed by phosphorylation and Ubl conjugation. Is it the reason why the image on the product page showing the band at 100 KDa?

    A: This product is a bit tricky as the protein is especially prone to forming protein complexes with other proteins; specifically 53/TP53, TP73/p73, RBL5 and RP11. Due to these interactions, it is common for this protein to be detected at a much higher weight than the expected weight given in Uniprot. In our testing, this protein usually detects a band at 100kDa. It is also possible to detect lower bands at 55 and 30kDa.

  • Q: I would like to know the epitope of the antibody HDM2 antibody SMP14 for human.

    A: We have not mapped the exact epitope for NB100-2736, but the immunogen is listed as a the synthetic peptide C-SRPSTSSRRRAISE, corresponding to amino acids 154-167 of human MDM2/HDM2. [UniProt# Q00987]

  • Q: I would like to know where I can expect to see the signals with NB100-2736 in WB. After checking the information from UniProt, the MW of MDM2 seems to be 55 KDa. However, it may be changed by phosphorylation and Ubl conjugation. Is it the reason why the image on the product page showing the band at 100 KDa?

    A: This product is a bit tricky as the protein is especially prone to forming protein complexes with other proteins; specifically 53/TP53, TP73/p73, RBL5 and RP11. Due to these interactions, it is common for this protein to be detected at a much higher weight than the expected weight given in Uniprot. In our testing, this protein usually detects a band at 100kDa. It is also possible to detect lower bands at 55 and 30kDa.

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