ENCAB000AMR

Antibody against Homo sapiens ZNF143

Homo sapiens
GM12878, HepG2, HeLa-S3, K562
characterized to standards
Homo sapiens
any cell type or tissue
partially characterized
Status
released
Source (vendor)
Proteintech
Product ID
16618-1-AP
Lot ID
8059
Characterized targets
ZNF143 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Antigen description
Raised against human ZNF143 (amino acids 283-626)-6his fusion protein.

Characterizations

ZNF143 (Homo sapiens)
Method: motif enrichment
compliant
Caption
The motif for target ZNF143 is represented by the attached position weight matrix (PWM) derived from ENCFF938XNY. Motif enrichment analysis was done by Dr. Zhizhuo Zhang (Broad Institute, Kellis Lab) using known motifs (http://compbio.mit.edu/encode-motifs/) and previously published ChIP-seq data (http://www.broadinstitute.org/~zzhang/motifpipeline/data/TrainSetInfo.txt). The accept probability score of the given transcription factor was calculated using a Bayesian approach. This analysis also includes three motif enrichment scores, computed by overlapping the motif instances with the given ChIP-seq peak locations, as well as an enrichment rank. For more information on the underlying statistical methods, please see the attached document. Accept probability score: 0.965199792 Global enrichment Z-score: 4.012241762 Positional bias Z-score: 3.954561752 Peak rank bias Z-score: 9.62442107 Enrichment rank: 1.0
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
ZNF143 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
not reviewed
Caption
Immunoprecipitation of ZNF143 from K562 cells using anti-ZNF143 (16618-1-AP) antibody. Lane 1: input nuclear lysate, Lane 2: material immunoprecipitated with 16618-1-AP, Lane 3: material immunoprecipitated using control IgG. Bands A, B and C were excised from the gel and subject to analysis by mass spectrometry. IP followed by masspectrometry: Briefly, protein was immunoprecipitated from K562 whole cell lysates using 16618-1-AP and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGE Bis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was silver-stained, gel fragments corresponding to the bands indicated were excised and destained using the SilverSNAP Stain for Mass Spectrometry (Pierce). Then proteins were trypsinized using the in-gel digestion method. Digested proteins were analyzed on an LTQ-Orbitrap (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Protein false discovery rate < 1%, 2 peptides per protein minimum). We report multiple proteins identified in band A, B and C, although many of these are also present in a control immunoprecipitation and are thus likely to present due to non-specific association with the IP matrix. Of the specifically immunoprecipitated proteins, ZNF143 is the enriched in all three bands. Based on these observations, this band is likely due to the presence of immunoprecipitated ZNF143 and 16618-1-AP meets the ENCODE standard for validation by this criterion.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
ZNF143 (Homo sapiens)
Method: immunoprecipitation
not reviewed
Caption
Western Blot analysis of nuclear extracts from GM12878, K562, HelaS3 and HepG2 using anti ZNF143 antibody from ProetinTech (16618-1-AP). Expected protein band 68kD. Immunoprecipitation: Western Blot analysis of immunoprecipitated proteins from HepG2 nuclear extracts detected using anti ZNF143 antibody from ProteinTech (16618-1-AP). Expected protein band 68kD. Western blot analysis using anti-ZNF143 (16618-1-AP) antibody detects a protein of ~68kD in multiple human cells. Immunoprecipitation using anti-ZNF143 (16618-1-AP) antibody significantly enriches a protein of ~68 kD.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
ZNF143 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Western Blot analysis of immunoprecipitated proteins from HepG2 nuclear extracts detected using anti-ZNF143 antibody from ProteinTech (16618-1-AP). Expected protein band size is 68 kD (as indicated by arrow).
Reviewer comment
Marked band is < 50% of the total signal in the lane.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
ZNF143 (Homo sapiens)
GM12878K562HeLa-S3HepG2
Method: immunoblot
Attachment from submitter
compliant
Caption
Western Blot analysis of nuclear extracts from GM12878, K562, HeLa-S3, and HepG2 using anti-ZNF143 antibody from ProteinTech (16618-1-AP). Expected protein band size is 68 kD (indicated by arrow)
Reviewer comment
Marked band in lane 4 not >50% of the total signal but ZNF143 detected in same smaller sized band in IP-MS in K562.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
ZNF143 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2 using the antibody 16618-1-AP. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 68.896.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
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