Anti-KMT6 / EZH2 antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab3748
• Prices: $403.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IHC-F WB IP ChIP IHC-P ICC/IF ICC/IF Radioimmunoprecipitation
• Species Reactivities: Mouse, Bovine, Human, Xenopus, Zebrafish
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human EZH2
• Blocking Peptide: KMT6 / EZH2 peptide
• Description: Rabbit Polyclonal
• Gene: EZH2
• Conjugate: Unconjugated

Product images

Anti-KMT6 / EZH2 antibody - ChIP Grade (ab3748) at 1 µg/ml + HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902) at 10 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Additional bands at : 23 kDa,48 kDa. We are unsure as to the identity of these extra bands.Exposure time : 16 minutes

All lanes : Anti-KMT6 / EZH2 antibody - ChIP Grade (ab3748) at 1/1000 dilutionLane 1 : HEK293 cell lysateLane 2 : Mouse hippocampus cell lysateLane 3 : Rat primary hippocampal neurons cell lysateLysates/proteins at 30 µg per lane.SecondaryHRP-conjugated Mouse anti-rabbit IgG at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Observed band size : 85 kDa (why is the actual band size different from the predicted?)Additional bands at : 100 kDa (possible non-specific binding),20 kDa (possible non-specific binding).Exposure time : 20 secondsThis image is courtesy of an anonymous AbreviewSee Abreview

IP with rabbit polyclonal to EZH2 (ab3748).U20S cells were transfected with 4ug of Myc-EZH2. WB reprobe using 0.4ug/ml in TBS milk 2%, BSA 0.5%.NB. ab3748 did not detect endogenous EZH2.

ab3748 (1/100) was used to stain EZH2 in mouse embronic stem cells. The antibody localised to the nucleus and colocalised with DNA. Some non-specific staining was observed in the cytoplasm. Cells were fixed for 15 min in 4% PFA. Blocking  was performed in PBS, 1% BSA, 0.1% TX-100. Hybridisation was in the same buffer at 4 degrees, o/n.

Anti-KMT6 / EZH2 antibody - ChIP Grade (ab3748) at 1 µg/ml + EZH2 - Recombinant Protein at 0.1 µgSecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.Observed band size : 40 kDa (why is the actual band size different from the predicted?)ab3748 detects a band at 40 kDa which corresponds to the predicted molecular weight of the Human EZH2 Recombinant Protein.

ab3748 staining KMT6 / EZH2 in human U-2 OS cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton ×100 and blocking with 1% BSA was done for 1 hour at RT. Samples were incubated with primary antibody (1/100) for 2 hours. An undiluted Alexa Fluor®594-conjugated goat polyclonal to rabbit IgG was used undiluted as secondary antibody.See Abreview

ab3748 staining KMT6 / EZH2 in murine hippocampus tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed using 10mM sodium citrate. Samples were then permeabilized using 0.3% Triton, blocked with 3% BSA for 30 minutes at 22°C and then incubated with ab3748 at a 1/3000 dilution for 16 hours at 4°C. The secondary used was a biotin conjugated horse polyclonal, used at a 1/2000 dilution. ABC method.See Abreview

ChIP analysis of Human glioma cells using ab3748 to bind KMT6 / EZH2. Chromatin was obtained by cross-linking with 1% formaldehyde for 10 minutes and incubated with primary antibody (1/250) for 16 hours at 4°C. Protein binding was detected using semi-quantitive PCR.See Abreview

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