YY1 Nuclear Loading Control Antibody (3P967) is a Rabbit antibody targeting YY1 Nuclear Loading Control. YY1 Nuclear Loading Control Antibody (3P967) can be used in FCM,IF,IHC-Fr,IHC-P,WB.

IgG

3P967

Human,Mouse,Rat

1. Paraformaldehyde-fixed, paraffin embedded (Human smooth muscle); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (mouse lymphoid); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (mouse ovary); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (mouse placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded (Rat lymphoid); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded (Rat bladder); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded (Rat ovary); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded (rat breast); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded (rat placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded (human breast carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (YY1 (Nuclear Loading Control)) Monoclonal Antibody, Unconjugated (TMAB-01992) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
13. Blank control: K562. Primary Antibody (green line): Rabbit Anti-YY1 (Nuclear Loading Control) antibody (TMAB-01992)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
14. Sample:
Lane 1: MCF-7 (Human) Cell Lysate at 30 μg
Lane 2: Jurkat (Human) Cell Lysate at 30 μg
Lane 3: Du145 (Human) Cell Lysate at 30 μg
Lane 4: U251 (Human) Cell Lysate at 30 μg
Lane 5: Panc-1 (Human) Cell Lysate at 30 μg
Lane 6: MDA-MB-231 (Human) Cell Lysate at 30 μg
Lane 7: Molt-4 (Human) Cell Lysate at 30 μg
Lane 8: Hela (Human) Cell Lysate at 30 μg
Lane 9: HL60 (Human) Cell Lysate at 30 μg
Lane 10: Urinary bladder (Mouse) Lysate at 40 μg
Lane 11: Testis (Mouse) Lysate at 40 μg
Primary: Anti-YY1 (TMAB-01992) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 65-68 kDa
Observed band size: 65 kDa

WB: 1:500-2000; IHC-P: 1:50-200; IHC-Fr: 1:50-200; IF: 1:50-200; FCM: 1ug/Test

Monoclonal

Recombinant Protein: human YY1

Human

YY1

Transcriptional repressor protein YY1

Zinc Finger,Other factors,Transcription Factors,YY1,Neurogenesis

Multifunctional transcription factor that exhibits positive and negative control on a large number of cellular and viral genes by binding to sites overlapping the transcription start site. Binds to the consensus sequence 5'-CCGCCATNTT-3'; some genes have been shown to contain a longer binding motif allowing enhanced binding; the initial CG dinucleotide can be methylated greatly reducing the binding affinity. The effect on transcription regulation is depending upon the context in which it binds and diverse mechanisms of action include direct activation or repression, indirect activation or repression via cofactor recruitment, or activation or repression by disruption of binding sites or conformational DNA changes. Its activity is regulated by transcription factors and cytoplasmic proteins that have been shown to abrogate or completely inhibit YY1-mediated activation or repression. For example, it acts as a repressor in absence of adenovirus E1A protein but as an activator in its presence. May play an important role in development and differentiation. Proposed to recruit the PRC2/EED-EZH2 complex to target genes that are transcriptional repressed. Involved in DNA repair. In vitro, binds to DNA recombination intermediate structures (Holliday junctions). Proposed core component of the chromatin remodeling INO80 complex which is involved in transcriptional regulation, DNA replication and probably DNA repair; proposed to target the INO80 complex to YY1-responsive elements.