ENCAB214KLU

Antibody against Homo sapiens ZNF800

Homo sapiens
HepG2, K562
characterized to standards with exemption
Status
released
Source (vendor)
Bethyl Labs
Product ID
A303-283A
Lot ID
1
Characterized targets
ZNF800 (Homo sapiens)
Host
rabbit
External resources

Characterizations

ZNF800 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
IP-Western Blot analysis of HepG2 whole cell lysate using ZNF800 specific antibody. Lane 1 is 1% of twenty million whole cell lysate input and lane 2 is 25% of IP enrichment using rabbit normal IgG (lanes under 'IgG'). Lane 3 is 1% of twenty million whole cell lysate input and lane 4 is 10% IP enrichment using rabbit polyclonal anti-ZNF800 antibody (lanes under 'ZNF800').
Submitter comment
The marked band is similar in size to that of the one seen in K562 that was analyzed by mass spec. We believe it is the protein of interest.
Reviewer comment
The expected size is 75kDa and the major immunoreactive band appears to be outside of the allowed 20% size deviation, but mass spec analysis in K562 of a similarly sized band revealed it to be the target of interest.
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD
ZNF800 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
Representative image of immunoprecipitation performed on whole cell extracts from the K562 cell line using the ZNF800-specific antibody A303-283A. Lane 1: Input from IP using control IgG. Lane 2: Immunoprecipitated material using control IgG. Lane 3: Input from IP using ZNF800 antibody. Lane 4: Immunoprecipitated material using ZNF800 antibody. Outlined regions were excised from gel and subjected to analysis by mass spectrometry. Target molecular weight: 75 kDa.
Submitter comment
The main immunoreactive band is higher than expected but we show by mass spectrometry that it is detecting the protein of interest.
Reviewer comment
Immunoreactive band is outside of allowed 20% deviation but accompanying mass spectrometry characterization shows it is the protein of interest.
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD
ZNF800 (Homo sapiens)
Method: knockdown or knockout
Attachment from submitter
not reviewed
Caption
Western blot following CRISPR against ZNF800 in K562 whole cell lysate using ZNF800 specific antibody. Lane 1 is a ladder, lane 2 is K562 non-targeting control knockdown, lane 3 and 4 are two different CRISPR against ZNF800.ZNF800 protein appears as the green band, Tubulin serves as a control and appears in red.
Reviewer comment
Acceptable levels of knockdown is seen for the marked immunoreactive signal, but not for the higher band that was confirmed by mass-spec.
Submitted by
Xintao Wei
Lab
Brenton Graveley, UConn
ZNF800 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
compliant
Caption
IP followed by mass spectrometry. Protein was immunoprecipitated from K562 whole cell lysates using the antibody A303-283A, loaded on a 4-12% NuPAGE Bis-Tris gel, and separated via electrophoresis. Using a reference western blot done in parallel, gel pieces corresponding to the sections indicated were excised and submitted for analysis by the UCSD Biomolecular and Proteomics Mass Spectrometry Facility.
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD