1. Antibodies
  2. Homo sapiens
  3. ZNF512B

ENCAB402XWG

Antibody against Homo sapiens ZNF512B

Homo sapiens
MCF-7
characterized to standards with exemption
Status
released
Source (vendor)
Bethyl Labs
Product ID
A303-235A
Lot ID
1
Characterized targets
ZNF512B (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1456
External resources

Characterizations

ZNF512B (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody A303-235A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 97.264
Reviewer comment
Multiple bands and the marked band is not 50% of the total signal in the lane. May be rescued with a siRNA knockdown.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
ZNF512B (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from MCF7 as rescue! nuclear cell lysates using the antibody A303-235A, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGEBis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was stained by ColloidialCoomassie G-250 stain, gel fragments corresponding to the bands indicated were excised. Then proteins were trypsinized using the in-gel digestion method. Digested proteins were analyzed on an Orbitrap Elite mass spectrometer (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Peptide false discovery rate < 1%, 2 unique peptides per protein minimum, mass error < 10 ppm).
Submitter comment
None of the proteins ranked above have been shown to be sequence-specific TFs (including SET1A and HCFC1).
Submitted by
Jessika Adrian
Lab
Michael Snyder, Stanford
ZNF512B (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line MCF7 using the antibody A303-235A. Lane 1: input nuclear lysate. Lane 2: material immunoprecipitated with antibody. Lane 3: material immunoprecipitated using control IgG. Marked bands were excised from gel and subjected to analysis by mass spectrometry. Target molecular weight: 97.264.
Submitter comment
We followed up the IP with mass-spec analysis to demonstrate the presence of the targeted TF in immunoreactive bands.
Reviewer comment
Multiple immunoreactive bands and band of expected size is not >50% of the total immunoreactive signal in the lane but the target TF is the highest rank sequence-specific factor in both analyzed bands in the accompanying mass-spec analysis (though finer grained slices would have been better).
Submitted by
Jessika Adrian
Lab
Michael Snyder, Stanford