Revision 6
#97800
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IF-IC, ChIP, ChIP-seq, C&R, C&T
Reactivity:
H M Mk
Sensitivity:
Endogenous
MW (kDa):
50-75
Source/Isotype:
Rabbit IgG
UniProt ID:
#O75030
Entrez-Gene Id:
4286
Product Usage Information
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunofluorescence (Immunocytochemistry) | 1:100 - 1:200 |
Chromatin IP | 1:50 |
Chromatin IP-seq | 1:50 |
CUT&RUN | 1:50 |
CUT&Tag | 1:50 |
Specificity/Sensitivity
MITF (D3B4T) Rabbit mAb recognizes endogenous levels of total MITF protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human MITF protein.
Background
Microphthalmia-associated transcription factor (MITF) is a basic helix-loop-helix leucine zipper transcription factor that is most widely known for its roles in melanocyte, ophthalmic, and osteoclast development (1-3). In humans, MITF can function as a melanoma oncogene (4) and mutations in the corresponding MITF gene are associated with Waardenburg syndrome type 2, an auditory-pigmentary syndrome characterized by developmental defects in cells derived from neural crest (5). At least 12 isoforms of MITF have been identified, which exhibit differential patterns of expression among cell and tissue types (6).
Background References
- Hershey, C.L. and Fisher, D.E. (2004) Bone 34, 689-96.
- Nomura, S. et al. (2001) J Bone Miner Metab 19, 183-7.
- Widlund, H.R. and Fisher, D.E. (2003) Oncogene 22, 3035-41.
- Levy, C. et al. (2006) Trends Mol Med 12, 406-14.
- Read, A.P. and Newton, V.E. (1997) J Med Genet 34, 656-65.
- Tachibana, M. (2000) Pigment Cell Res 13, 230-40.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP C&R: CUT&RUN C&T: CUT&Tag
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 6
#97800
MITF (D3B4T) Rabbit mAb
Western blot analysis of extracts from various cell lines using MITF (D3B4T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Confocal immunofluorescent analysis of SK-MEL-28 cells (left, positive) and HCT 116 cells (right, negative) using MITF (D3B4T) Rabbit mAb (green), DyLight™ 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Chromatin immunoprecipitations were performed with cross-linked chromatin from SK-MEL-28 cells and MITF (D3B4T) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across BEST1/VMD2, a known target gene of MITF (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Revision 6
#97800
MITF (D3B4T) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from SK-MEL-28 cells and 10 μl of MITF (D3B4T) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 11 (upper), including BEST1/VMD2 (lower), a known target gene of MITF (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from SK-MEL-28 cells and either MITF (D3B4T) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human Tyrosinase Promoter Primers #12970, human VMD2 intron 1 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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.
CUT&RUN was performed with SM-MEL-28 cells and MITF (D3B4T) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across TYR, a known target gene of MITF (see additional figure containing CUT&RUN-qPCR data).
Revision 6
#97800
MITF (D3B4T) Rabbit mAb
CUT&RUN was performed with SM-MEL-28 cells and MITF (D3B4T) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 11 (upper), including TYR (lower), a known target gene of MITF (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with SM-MEL-28 cells and either MITF (D3B4T) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human Tyrosinase Promoter Primers #12970 and SimpleChIP® Human α Satellite Repeat Primers #4486. 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.
CUT&Tag was performed with SK-MEL-28 cells and MITF (D3B4T) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across BEST1/VMD2, a known target gene of MITF (see our ChIP-qPCR figure).
Revision 6
#97800
MITF (D3B4T) Rabbit mAb
CUT&Tag was performed with SK-MEL-28 cells and MITF (D3B4T) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 11 (upper), including BEST1/VMD2 (lower), a known target gene of MITF (see our ChIP-qPCR figure).