ENCAB214KLU
Antibody against Homo sapiens ZNF800
Homo sapiens
K562, HepG2
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
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
- Grant
- U54HG007005
ZNF800 (Homo sapiens)
K562
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
- Grant
- U54HG007005
- Download
- K562_Bethyl_A303-283A_1_ZNF800.png
ZNF800 (Homo sapiens)
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
- Grant
- U54HG007005
- Download
- updated_ZNF800-CRISPR-K562.png
ZNF800 (Homo sapiens)
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
- Grant
- U54HG007005
- Download
- ZNF800 K562 MassSpec.pdf