MITF antibody (mAb)

RRID: AB_2614955
Clone: C5.D5
Aliases: MI, WS2, CMM8, WS2A, bHLHe32
Catalog No: 39789 Format: 100 µg $455 Buy
Catalog No: 39790 Format: 10 µg $110 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG1
Purification:
Protein G Chromatography
Host:
Mouse
Molecular Weight:
52-56 kDa
Reactivity:
Human

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunoprecipitation Validated Immunofluorescence Validated Electrophoretic Mobility Shift Assay Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 2.5 - 10 µg per ChIP
ChIP-Seq: 4 µg each

This antibody is also available as an AbFlex® engineered recombinant antibody. For details on the corresponding AbFlex Recombinant Antibody, see Catalog No. 91201.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

IP
WB
ICC/IF
EMSA

Please see the original reference for clone C5, which has been characterized for IP, WB, ICC/IF, and EMSA.

Immunogen

This MITF antibody was raised against an N-terminal fragment of human MITF.

Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

References

 

MITF antibody (mAb) tested by ChIP-Seq.
(ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) and chromatin from a human melanoma cell line (2.5 million cells). ChIP DNA was sequenced on the Illumina GA II and 19 million sequence tags were mapped to identify MITF binding sites across the genome. The image shows hundreds of MITF binding sites across a 60 million bp region on chromosome 9.

MITF antibody (mAb) tested by ChIP.
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of chromatin from human lung melanoma cell line MALME-3M and 4 µg of MITF antibody. ChIP DNA was used in qPCR with the control primer pairs or gene-specific primer pairs as indicated. Data are presented as Binding Events Detected per 1000 Cells using Active Motif's Epigenetic Services normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.

Background

MITF protein (Microphthalmia-associated Transcription Factor) is a basic helix-loop-helix type transcription factor involved in the differentiation and development of retinal pigment epithelium and melanocytes. It is also responsible for pigment cell-specific transcription of the melanogenesis enzyme genes tyrosinase and tyrosinase related protein 1. For activity MITF needs to heterodimerize with other bHLH proteins such as TFE3, TFEB or TFE. Defects in MITF are the cause of Waardenburg syndrome type 2A and Tietz syndrome.

qPCR Primers

Active Motif's Human Negative Control Primer Set 2 (Cat. No. 71002) has been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples. For ChIP experiments with this antibody on mouse samples, the Mouse Positive Control Primer Set Pax-2 (Cat. No. 71020) and Mouse Negative Control Primer Set 3 (Cat. No. 71013) have been validated for both qPCR and endpoint PCR.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with MITF antibody (mAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • WB = Western Blotting