Rabbit Recombinant Monoclonal RUNX1 / AML1 antibody. Suitable for ChIC/CUT&RUN-seq, IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 5 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
ChIC/CUT&RUN-seq | IP | ChIP | WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|---|---|---|
Human | Tested | Tested | Not recommended | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info 5 µg | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/2000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Forms the heterodimeric complex core-binding factor (CBF) with CBFB. RUNX members modulate the transcription of their target genes through recognizing the core consensus binding sequence 5'-TGTGGT-3', or very rarely, 5'-TGCGGT-3', within their regulatory regions via their runt domain, while CBFB is a non-DNA-binding regulatory subunit that allosterically enhances the sequence-specific DNA-binding capacity of RUNX. The heterodimers bind to the core site of a number of enhancers and promoters, including murine leukemia virus, polyomavirus enhancer, T-cell receptor enhancers, LCK, IL3 and GM-CSF promoters (Probable). Essential for the development of normal hematopoiesis (PubMed:17431401). Acts synergistically with ELF4 to transactivate the IL-3 promoter and with ELF2 to transactivate the BLK promoter (PubMed:10207087, PubMed:14970218). Inhibits KAT6B-dependent transcriptional activation (By similarity). Involved in lineage commitment of immature T cell precursors. CBF complexes repress ZBTB7B transcription factor during cytotoxic (CD8+) T cell development. They bind to RUNX-binding sequence within the ZBTB7B locus acting as transcriptional silencer and allowing for cytotoxic T cell differentiation. CBF complexes binding to the transcriptional silencer is essential for recruitment of nuclear protein complexes that catalyze epigenetic modifications to establish epigenetic ZBTB7B silencing (By similarity). Controls the anergy and suppressive function of regulatory T-cells (Treg) by associating with FOXP3. Activates the expression of IL2 and IFNG and down-regulates the expression of TNFRSF18, IL2RA and CTLA4, in conventional T-cells (PubMed:17377532). Positively regulates the expression of RORC in T-helper 17 cells (By similarity). Isoform AML-1G shows higher binding activities for target genes and binds TCR-beta-E2 and RAG-1 target site with threefold higher affinity than other isoforms. It is less effective in the context of neutrophil terminal differentiation. Isoform AML-1L interferes with the transactivation activity of RUNX1.
AML1, CBFA2, RUNX1, Runt-related transcription factor 1, Acute myeloid leukemia 1 protein, Core-binding factor subunit alpha-2, Oncogene AML-1, Polyomavirus enhancer-binding protein 2 alpha B subunit, SL3-3 enhancer factor 1 alpha B subunit, SL3/AKV core-binding factor alpha B subunit, CBF-alpha-2, PEA2-alpha B, PEBP2-alpha B
Rabbit Recombinant Monoclonal RUNX1 / AML1 antibody. Suitable for ChIC/CUT&RUN-seq, IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 5 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
RUNX1 also known as AML1 is a transcription factor with a molecular weight of approximately 48 kDa. It belongs to the Runt-related transcription factor family and plays a critical role in hematopoiesis. RUNX1 is expressed in hematopoietic stem cells and various other tissues where it regulates the expression of genes involved in the differentiation and proliferation of blood cells. It exerts its function by binding to specific DNA sequences thereby controlling the transcriptional activity necessary for normal hematopoietic development.
RUNX1 is essential in the formation of blood cells and is part of the core-binding factor (CBF) complex. This complex is a heterodimer comprising RUNX1 and the CBFβ subunit. The interaction between RUNX1 and CBFβ stabilizes the DNA binding capability of RUNX1 facilitating the activation of target gene transcription. The proper functioning of RUNX1 is necessary for the maintenance of normal lineage specification of hematopoietic progenitors affecting both myeloid and lymphoid cell lineages.
RUNX1 plays a significant role in the Wnt signaling pathway and the TGF-beta signaling pathway. RUNX1 interacts with several proteins in these pathways including SMAD proteins and β-catenin which are important for transmitting extracellular signals that regulate cell growth and differentiation. RUNX1’s role in these pathways highlights its importance not only in hematopoiesis but also in preventing abnormal cell proliferation.
RUNX1 mutations are strongly associated with acute myeloid leukemia (AML) and familial platelet disorder. In AML RUNX1 mutations disrupt normal hematopoiesis leading to the uncontrolled proliferation of immature blood cells. RUNX1-related proteins such as the GM-CSF receptor can contribute to disease progression by altering cytokine signaling. RUNX1's involvement in familial platelet disorder reflects its importance in maintaining normal blood cell counts and function as loss of RUNX1 function leads to predisposition to leukemia.
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Terms & Conditions.
RUNX1 / AML1 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10ug with ab240639 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution. Lane 1: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10ug
Lane 2: ab240639 IP in Jurkat whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab240639 in Jurkat whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds
All lanes: Immunoprecipitation - Anti-RUNX1 / AML1 antibody [EPR23044-100] (ab240639)
Predicted band size: 48 kDa
Blocking and diluting buffer and concentration: 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 23352661, 29296779). The RUNX1 gene has several isoforms, 3 major isotypes. RUNX1b is broadly expressed, and RUNX1a overexpression has been reported in AML.
Exposure time: Lane 1: 48 seconds Lanes 2-3: 26 seconds
All lanes: Western blot - Anti-RUNX1 / AML1 antibody [EPR23044-100] (ab240639) at 1/1000 dilution
Lane 1: THP-1 (human monocytic leukemia monocyte), whole cell lysate at 10 µg
Lane 2: Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 10 µg
Lane 3: MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 27-55 kDa
Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling RUNX1 / AML1 with ab240639 at 1/2000 dilution (0.25 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on human breast tissue is observed. The section was incubated with ab240639 for 15 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling RUNX1 / AML1 with ab240639 at 1/2000 dilution (0.25 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on human breast carcinoma (PMID: 24967588) tissue is observed. The section was incubated with ab240639 for 15 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat (Human T cell leukemia T lymphocyte) cells labelling RUNX1 / AML1 with ab240639 at 1/100 dilution, followed by ab240639 anti-RUNX1/AML1 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing strong nuclear and weak cytoplasmic staining in Jurkat cell line is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab240639 anti-RUNX1/AML1 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) cells labelling RUNX1 / AML1 with ab240639 at 1/500 (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730, Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 K-562 (Human chronic myelogenous leukemia lymphoblast) cells and 5µg of ab240639 [EPR23044-100]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.
Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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