WDR5 Polyclonal Antibody

$HeLa cells were stained with the Bioss antibody against WDR5 (Cat. No. bs-53101R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the WDR5 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.$

HeLa cells were stained with the Bioss antibody against WDR5 (Cat. No. bs-53101R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the WDR5 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

$Nuclear extracts from HeLa cells (20 \u03bcg) were analyzed by Western blot using the Bioss antibody against WDR5 (Cat. No. bs-53101R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.$

Nuclear extracts from HeLa cells (20 \u03bcg) were analyzed by Western blot using the Bioss antibody against WDR5 (Cat. No. bs-53101R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

$ChIP was performed on sheared chromatin from 4 million HeLa cells using 2 \u03bcg of the Bioss antibody against WDR5 (Cat. No. bs-53101R) as described above. The IP\u2019d DNA was subsequently analyzed on an Illumina HiSeq. Library preparation, cluster generation, and sequencing were performed according to the manufacturer\u2019s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. The figure shows the enrichment along the complete sequence and a 2 Mb region of human chromosome 1 (fig A and B), and in two genomic regions surrounding the RPL41 and RPS4X positive control genes (fig C and D).$

ChIP was performed on sheared chromatin from 4 million HeLa cells using 2 \u03bcg of the Bioss antibody against WDR5 (Cat. No. bs-53101R) as described above. The IP\u2019d DNA was subsequently analyzed on an Illumina HiSeq. Library preparation, cluster generation, and sequencing were performed according to the manufacturer\u2019s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. The figure shows the enrichment along the complete sequence and a 2 Mb region of human chromosome 1 (fig A and B), and in two genomic regions surrounding the RPL41 and RPS4X positive control genes (fig C and D).

$ChIP assays were performed using HeLa cells, the Bioss antibody against WDR5 (Cat. No. bs-53101R) and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 4 million cells. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the RPL41 and RPS4X ribosomal protein genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

ChIP assays were performed using HeLa cells, the Bioss antibody against WDR5 (Cat. No. bs-53101R) and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 4 million cells. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the RPL41 and RPS4X ribosomal protein genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Applications

WB
IF
ChIP-seq

Reactivity

Human

Overview
Catalog #	bs-53101R
Product Name	WDR5 Polyclonal Antibody
Applications	WB IF CHIP-SEQ
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Polyclonal antibody raised in rabbit against human WDR5 (WD (tryptophan-aspartate) repeat domain 5), using a recombinant protein
Clonality	Polyclonal
Isotype	Not Determined
Concentration	2ug/ul
Purification	Protein G purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Store at -20C for 12 months.
Target
Gene ID	11091
Swiss Prot	P61964
Synonyms	WD repeat-containing protein 5, WDR5, BMP2-induced 3-kb gene protein, BIG3
Background	Contributes to histone modification. May position the N-terminus of histone H3 for efficient trimethylation at 'Lys-4'. As part of the MLL1/MLL complex it is involved in methylation and dimethylation at 'Lys-4' of histone H3. H3 'Lys-4' methylation represents a specific tag for epigenetic transcriptional activation. As part of the NSL complex it may be involved in acetylation of nucleosomal histone H4 on several lysine residues. May regulate osteoblasts differentiation.
Application Dilution
WB	1:300-5000
IF
ChIP-seq