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c-Myc Recombinant Rabbit Monoclonal Antibody [PS00-11]

Usd: 385

Specification

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Catalog# HA721182

c-Myc Recombinant Rabbit Monoclonal Antibody [PS00-11]

Application

  • WB

  • IHC-P

  • IP

  • ChIP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

c-Myc Recombinant Rabbit Monoclonal Antibody [PS00-11]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within N terminus of Human c-Myc.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP, ChIP

Molecular Weight

Predicted band size: 49 kDa

Positive Control

HeLa cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, C6 cell lysate, HEK-293 cell lysate, Jurkat cell lysate, K-562 cell lysate, human B lymphocytoma tissue, human colon tissue, human colon carcinoma tissue.

Conjugation

unconjugated

Clone Number

PS00-11

RRID

AB_3072302

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:200

  • IP

  • 1-2μg/sample

  • ChIP

  • Use 0.5~2 μg for 25 μg of chromatin.

Target

Function

The proto-oncogene c-MYC is located on chromosome 8q24. It encodes a nuclear posphoprotein that acts as a growth promotor and transcription factor regulating a variety of cellular functions such as cell growth, differentiation and apoptosis. c-MYC is involved in the regulation of 10-15% of all human genes and is estimated to be involved in 20% of all human cancers. c-MYC is essential for early B-cell development in the bone marrow and the unregulated expression of c-MYC in germinal center B-cells increases the chances of oncogentic events for the development of B-cell lymphomas. Mutations, amplification, rearrangement, and translocation of the c-MyC gene have been associated with various hematopoietic tumors, leukemias, and lymphomas as well as a variety of carcinomas (e.g. thyroid, breast, endometrium) and gliomas. IHC for c-MYC overexpression (>40%) is used to screen and predict c-MYC rearrangements and distinguish diffuse large B-cell lymphoma (DLBCL) NOS from high grade B-cell lymphoma with c-MYC, BCL2 and/or BCL6 rearrangements or so-called “double or triple hit lymphomas” who tend to have aggressive behavior and poor outcome when treated with standard therapy. c-MYC protein is overexpressed in 90% of Burkitt lymphomas (BL) and caused by c-Myc gene translocation. tonsil and colon are recommended as positive and negative tissue controls for c-MYC. In the tonsil, protocols must be calibrated to provide a moderate to strong, distinct nuclear staining reaction in approximately 10% of lymphocytes scattered both in the interfollicular zones and in the reactive germinal centers of the tonsil. A weak, distinct nuclear staining reaction of mantle zone B-cells (app. 10-20%) should be seen. In colon, a weak to moderate nuclear staining reaction should be displayed in scattered epithelial cells in the basal crypts, whereas the luminal epithelial cells and smooth muscle cells of the tunica muscularis should be unstained.

Background References

1. Wu H. et. al. Tumor Necrosis Factor Receptor-Associated Factor 6 Promotes Hepatocarcinogenesis by Interacting With Histone Deacetylase 3 to Enhance c-Myc Gene Expression and Protein Stability. Hepatology. 2020 Jan

2. Tsounis D. et. al. C-myc oncogene activation in pharyngeal-esophageal squamous cell carcinoma. J BUON. 2021 Jul-Aug

Subcellular Location

Nucleoplasm, nucleolus

UNIPROT

SwissProt: P01106 Human

SwissProt: P01108 Mouse

SwissProt: P09416 Rat

Synonyms

AU016757 antibody

Avian myelocytomatosis viral oncogene homolog antibody

bHLHe39 antibody

c Myc antibody

Cellular myelocytomatosis oncogene antibody

Class E basic helix-loop-helix protein 39 antibody

MGC105490 antibody

MRTL antibody

Myc antibody

Myc protein antibody

Expand

Images

  • Western blot analysis of c-Myc on different lysates with Rabbit anti-c-Myc antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Neuro-2a cell lysate<br />Lane 3: NIH/3T3 cell lysate<br />Lane 4: C6 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 49 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 1 minute; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of c-Myc on different lysates with Rabbit anti-c-Myc antibody (HA721182) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Neuro-2a cell lysate
    Lane 3: NIH/3T3 cell lysate
    Lane 4: C6 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 49 kDa
    Observed band size: 55 kDa

    Exposure time: 1 minute; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721182) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of c-Myc on different lysates with Rabbit anti-c-Myc antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/1,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate<br />Lane 2: HeLa cell lysate<br />Lane 3: Jurkat cell lysate<br />Lane 4: K-562 cell lysate<br />Lane 5: Neuro-2a cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 49 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 1 minute; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of c-Myc on different lysates with Rabbit anti-c-Myc antibody (HA721182) at 1/1,000 dilution.

    Lane 1: HEK-293 cell lysate
    Lane 2: HeLa cell lysate
    Lane 3: Jurkat cell lysate
    Lane 4: K-562 cell lysate
    Lane 5: Neuro-2a cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 49 kDa
    Observed band size: 55 kDa

    Exposure time: 1 minute; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721182) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human B lymphocytoma tissue with Rabbit anti-c-Myc antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human B lymphocytoma tissue with Rabbit anti-c-Myc antibody (HA721182) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721182) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-c-Myc antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-c-Myc antibody (HA721182) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721182) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-c-Myc antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-c-Myc antibody (HA721182) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721182) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • c-Myc was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (<a href="/products/NBI01H" style="font-weight: bold;text-decoration: underline;">NBI01H</a>) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 30 seconds; ECL: K1801

    c-Myc was immunoprecipitated from 0.2 mg HeLa cell lysate with HA721182 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA721182 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA721182 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA721182 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 30 seconds; ECL: K1801

  • Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with c-Myc (<a href="/products/HA721182" style="font-weight: bold;text-decoration: underline;">HA721182</a>) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with c-Myc (HA721182) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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