Western blot analysis was performed using anti-MLL1 (Abcam) and anti-β-actin (Santa Cruz, CA, USA) antibodies. Cell lysates were prepared using radioimmunoprecipitation assay buffer (RIPA) (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 2 mM ethylenediaminetetraacetic acid, 1% sodium dodecyl sulfate (SDS), 1% sodium deoxycholate, and 1% NP- 40). Total cell lysates were ultrasonically dispersed for 10 min and centrifuged at 10,000× g for 20 min at 4 °C. The proteins were transferred to Immun-Blot polyvinylidene difluoride (PVDF) membranes (Bio-Rad, Hercules, CA, USA). Primary antibodies were used at a dilution of 1:1000 and then further incubated at 4 °C overnight. The membranes were then incubated with horseradish peroxidase (HRP)-labeled anti-rabbit second antibodies for 2 h at 1:5000 diluting in blocking solution. Expression levels were quantified by densitometry, followed by normalization to the β-actin.
Anti mll1
Manufactured by Abcam
Sourced in United States
Anti-MLL1 is a primary antibody that recognizes the MLL1 (Mixed Lineage Leukemia 1) protein. MLL1 is a histone methyltransferase that plays a crucial role in regulating gene expression during development and hematopoiesis. This antibody can be used for various research applications, such as Western blotting and immunoprecipitation, to detect and study the MLL1 protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Immun blot polyvinylidene difluoride pvdf membrane, by Bio-Rad (1 mentions)
Anti h4k16ac, by Merck Group (1 mentions)
Anti ar, by Thermo Fisher Scientific (1 mentions)
Anti ash2l, by Abcam (1 mentions)
Anti h3k4me3, by Abcam (1 mentions)
Anti h3k27me3, by Abcam (1 mentions)
Jetprime transfection reagent, by Polyplus Transfection (1 mentions)
Anti jmjd3, by Merck Group (1 mentions)
Chip kit, by Merck Group (1 mentions)
Anti h3k27me3 and, by Merck Group (1 mentions)
Anti mll2, by Abcam (1 mentions)
Rotor gene 3000, by Qiagen (1 mentions)
3 protocols using anti mll1
1
Western Blot Analysis of MLL1 Protein
2
ChIP Assay for AR and Chromatin Modifications
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ChIP assays were performed according to previously described protocols (25 (link)). 22Rv1 cells were transfected with FLAG-BAP18 using jetPRIME transfection reagent (Polyplus) or infected with shBAP18, and then cultured for 2 days in phenol red-free RPMI 1640 supplemented with 10% charcoal-dextran-stripped FBS. At ∼90% confluency, cells were treated with 10−8M DHT or EtOH for 12 h and harvested for ChIP. Immunoprecipitation of sonicated chromatin solutions was conducted by overnight incubation at 4°C with anti-AR (Thermo), anti-BAP18, anti-MLL1, anti-hMOF, anti-Ash2L, anti-H3K27me3 (Abcam), anti-H3K4me3 (Abcam) or anti-H4K16ac (Millipore) antibody. Cross-linking was reversed at 65°C, and DNA fragments were extracted with phenol-chloroform and precipitated with ethanol. The purified DNA was dissolved in TE buffer and analyzed by regular PCR. Primer sequences of PSA-ARE I/II were as follows: forward 5′-GCCAAGACATCTATTTCAGGAGC-3′, and reverse 5′-CCCACACCCAGAGCTGTGGAAGG-3′. DNA fragments was analyzed by real-time PCR (RT-PCR) with SYBR Green dye. Results were expressed as percentage of input chromatin and were derived from a single experiment that is representative of at least two independent experiments.
3
ChIP Assay for Histone Modifications
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The ChIP assay was performed with a ChIP kit (Millipore), as described previously [26 (link)]. Anti-H3K27me3 and anti-H3K4me3 antibodies were purchased from Millipore; anti-JMJD3, anti-MLL1, anti-MLL2, anti-MLL3, and anti-MLL4 antibodies were provided by Abcam. The immunoprecipitated DNA was purified, then subjected to quantitative polymerase chain reaction (qPCR) analysis using a Rotor-Gene 3000 thermocycler (Corbett Research Ltd.), with input DNA (total chromatin) as an endogenous control. The primers of HPK1 promoter were as follows: 5′-TGGGGAGATAGAGGTTGCAG-3′ (forward) and 5′-CGCCAGAAATCCAATGACTT-3′ (reverse).
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