Rabbit Recombinant Monoclonal SUZ12 antibody. Carrier free. Suitable for ChIC/CUT&RUN-seq, IP, ChIP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
ChIC/CUT&RUN-seq | IP | ChIP | WB | IHC-P | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Not recommended | Expected | Expected |
Mouse | Predicted | Predicted | Predicted | Predicted | Not recommended | Expected | Expected |
Rat | Predicted | Predicted | Predicted | Predicted | Not recommended | Predicted | Predicted |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Human, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Rat | Dilution info - | Notes - |
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Polycomb group (PcG) protein. Component of the PRC2 complex, which methylates 'Lys-9' (H3K9me) and 'Lys-27' (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene (PubMed:15225548, PubMed:15231737, PubMed:15385962, PubMed:16618801, PubMed:17344414, PubMed:18285464, PubMed:28229514, PubMed:29499137, PubMed:31959557). The PRC2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems (PubMed:12351676, PubMed:12435631, PubMed:15099518, PubMed:15225548, PubMed:15385962, PubMed:15684044, PubMed:16431907, PubMed:18086877, PubMed:18285464). Genes repressed by the PRC2 complex include HOXC8, HOXA9, MYT1 and CDKN2A (PubMed:15231737, PubMed:16618801, PubMed:17200670, PubMed:31959557).
CHET9, JJAZ1, KIAA0160, SUZ12, KIAA0160, JJAZ1, CHET9, Polycomb protein SUZ12, Chromatin precipitated E2F target 9 protein, Joined to JAZF1 protein, Suppressor of zeste 12 protein homolog, ChET 9 protein
Rabbit Recombinant Monoclonal SUZ12 antibody. Carrier free. Suitable for ChIC/CUT&RUN-seq, IP, ChIP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
Yes
EPR5234(N)
Affinity purification Protein A
Blue Ice
+4°C
+4°C
Do Not Freeze
ab249844 is the carrier-free version of Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Chromatin was prepared from HeLa cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 μg of chromatin, 5 μg of Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 (red), and 20 μl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187).
Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 (purified) at 1/20 dilution (16 μg/mL) immunoprecipitating SUZ12 in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg.
Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg
Lane 2 (+): Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 in HeLa whole cell lysate
For western blotting, Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 at 1/500 dilution (0.636 μg/mL) and veriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution was used.
Blocking and diluting buffer: 5% NFDM /TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187).
All lanes: Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187)
Predicted band size: 83 kDa
This data was developed using the same antibody clone in a different buffer formulation (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187).
Lanes 1 - 2: Merged signal (red and green). Green - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 observed at 90 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 was shown to react with SUZ12 in wild-type HAP1 cells in Western blot with loss of signal observed in SUZ12 knockout sample. Wild-type HAP1 and SUZ12 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187) at 1/1000 dilution
Lane 1: Wild-type HAP1 cell lysate at 20 µg
Lane 2: SUZ12 knockout HAP1 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 90 kDa
Chromatin was prepared from F9 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci).
Primers and probes are located in the first kb of the transcribed region.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187).
This data was developed using the same antibody clone in a different buffer formulation (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187).
Lanes 1- 2: Merged signal (red and green). Green - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 was shown to react with SUZ12 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line Human SUZ12 knockout HeLa cell line ab264983 (CRISPR/Cas9 edited cell lysate Human SUZ12 knockout HeLa cell lysate ab257721) lane below 100kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and SUZ12 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: SUZ12 CRISPR/Cas9 edited HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 100 kDa
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 5µg of Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 [EPR5234(N)]. 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.
This data was developed using the same antibody clone in a different buffer formulation (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187).
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