Overview

  • Product name
    Anti-Mel18 antibody - ChIP Grade
    See all Mel18 primary antibodies
  • Description
    Goat polyclonal to Mel18 - ChIP Grade
  • Host species
    Goat
  • Tested applications
    Suitable for: IP, ICC/IF, ChIP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide: KMTVNGAPVPPLT, corresponding to C terminal amino acids 332-344 of Human Mel18.

  • Positive control
    • Human lung lysate.
  • General notes
    Gene Ontology terms - transcription factor activity; regulation of transcription, DNA-dependent; nucleus. GenBank Accession Number – NP_009075.

Properties

Applications

Our Abpromise guarantee covers the use of ab5267 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP Use at an assay dependent dilution. PubMed: 19636380
ICC/IF 1/100.
ChIP Use at an assay dependent concentration.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).Can be blocked with Human Mel18 peptide (ab23085).

Target

  • Function
    Transcriptional repressor. Binds specifically to the DNA sequence 5'-GACTNGACT-3'. Has tumor suppressor activity. May play a role in control of cell proliferation and/or neural cell development. Regulates proliferation of early T progenitor cells by maintaining expression of HES1. Also plays a role in antero-posterior specification of the axial skeleton and negative regulation of the self-renewal activity of hematopoietic stem cells. Component of the Polycomb group (PcG) multiprotein PRC1 complex, a complex required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility.
  • Tissue specificity
    Detected in all tissues examined with high expression found in placenta lung and kidney and low expression, in liver, pancreas and skeletal muscle.
  • Sequence similarities
    Contains 1 RING-type zinc finger.
  • Post-translational
    modifications
    Phosphorylated. Homodimer formation is regulated by phosphorylation state with unphosphorylated protein forming homodimers.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • DNA binding protein Mel 18 antibody
    • DNA-binding protein Mel-18 antibody
    • Mel 18 antibody
    • MGC10545 antibody
    • PCGF 2 antibody
    • PCGF2 antibody
    • PCGF2_HUMAN antibody
    • Polycomb group ring finger 2 antibody
    • Polycomb group RING finger protein 2 antibody
    • RING finger protein 110 antibody
    • RNF110 antibody
    • Zinc finger protein 144 antibody
    • ZNF 144 antibody
    • ZNF144 antibody
    see all

Images

  • ab5267 staining (1µg/ml) of Human Lung lysate (RIPA buffer, 30µg total protein per lane).  Primary incubated for 1 hour.  Detected by western blot using chemiluminescence.

    ab5267 staining (1µg/ml) of Human Lung lysate (RIPA buffer, 30µg total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence.

References

This product has been referenced in:
  • Klauke K  et al. Polycomb Cbx family members mediate the balance between haematopoietic stem cell self-renewal and differentiation. Nat Cell Biol 15:353-62 (2013). WB . Read more (PubMed: 23502315) »
  • Holdt LM  et al. Alu elements in ANRIL non-coding RNA at chromosome 9p21 modulate atherogenic cell functions through trans-regulation of gene networks. PLoS Genet 9:e1003588 (2013). Read more (PubMed: 23861667) »
See all 12 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - whole cell (NT2 cell line)
Negative control
Intergenic region on Chromosome 11.
Specification
NT2 cell line
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Positive control
CCND2 and ALX4 genes

Tatiana Kan

Verified customer

Submitted Oct 17 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (NB4 cell line(acute promylocytic leukemia))
Specification
NB4 cell line(acute promylocytic leukemia)
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Mr. Sungsin Jo

Verified customer

Submitted Oct 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - whole cell (HL60(acute promyelotic leuemia cells line))
Specification
HL60(acute promyelotic leuemia cells line)
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: SDS lysis buffer
Detection step
Semiquantitative PCR
Positive control
acetly-histone H3 (cat #06-599, millipore)
Negative control
Rabbit IgG(cat# pp64, millipore)

Mr. Sungsin Jo

Verified customer

Submitted Oct 23 2009

Answer

Thank you for your enquiry. I have consulted the EXPASY databases and I retrieved the following information: Mel18 - Tissue specificity: Detected in all tissues examined with high expression found in placenta, lung and kidney and low expression, in liver, pancreas and skeletal muscle. Ring1B - No tissue specificity information. The recommended positive control for Mel18 is human lung lysate. For Ring1B we do not have a recommended positive control. However, a reviewer has successfully used this antiserum in western blotting using mouse cultured cells (3T3) and mouse brain. This may be a useful alternative to HeLa cells as I cannot guarantee that these cells express Ring1B. I have obtained the following details of how the positive control to how Mel18 was prepared and run by western blotting. Western Blot: Approx 40 kDa band observed in Human Lung extracts (predicted MW of 40kDa according to NP_009075). Recommended for use at 1-2µg/ml. Tissue Lysis. Tissue chunks were weighed and cut into approx 1mm cubes using a razor blade. The tissue was transferred to a handheld homogenizer and 3 ml of ice-cold RIPA buffer was added per 1g of tissue. The tissue was gently homogenised over 20 minutes on ice. The resulting lysate was aliquotted into 1.5 ml microfuge tubes and centrifuged at 13,000 rpm for 5 min in a microfuge. The supernatant was transferred into clean tubes and its protein concentration was measured with BioRad protein assay. The concentration was then adjusted to 5 mg/ml with RIPA lysis buffer. An equal volume of 2 x SDS sample buffer was added and the cell lysate was boiled for 5 minutes. Lysates were stored at -80C until use.(RIPA buffer = 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM PMSF, 1 mM EDTA, 5 µg/ml Aprotinin, 5 µg/ml Leupeptin, 1% Triton X-100, 1% Sodium deoxycholate, 0.1% SDS). SDS PAGE. Samples were run at 200V constant on a 12% acrylamide SDS-PAGE mini gel - using Biorad Mini-Protean 3 kit and protocols. Before loading samples had 5% (v/v) 2-ME added and were boiled for 3 minutes. Transfer. We used a Biorad Mini Trans-Blot, constant 100 V for 1 hour. Transfer Buffer was 20 mM Tris pH 8.0, 150 mM Glycine, 10% Methanol. We transferred to Millipore PVDF membrane and stained with Ponceau Red to evaluate the transfer. Staining. The membrane was blocked in 2.5% skimmed milk in TBS-T (TBS + 0.05% Tween-20) for 1 hr at room temperature with agitation. Primary antibody was incubated for 1 hr at room temperature with agitation. We used [a competitor's] secondary 1:3000) for 1 hr at room temperature with agitation. We washed with TBST three times after primary and secondary antibody, each wash lasting for 5-10 mins. ECL-plus (Amersham) was used rather than ECL, which is considerably more sensitive. Final detection was on autoradiography film. I hope this information helps. Please do not hesitate to contact me should you require further assistance.

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