ENCAB292USO

Antibody against Homo sapiens ZBTB33

Homo sapiens
MCF-7, K562, HepG2, GM12878
characterized to standards
Homo sapiens
HEK293T
not characterized to standards
Status
released
Source (vendor)
Sigma
Product ID
HPA005732
Lot ID
R04529
Characterized targets
ZBTB33 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Isotype
IgG
Antigen description
Transcriptional regulator Kaiso recombinant protein epitope signature tag (PrEST)
Antigen sequence
SSSPDSAVSNTSLVPQADTSQNTSFDGSLIQKMQIPTLLQEPLSNSLKISDIITRNTNDPGVGSKHLMEGQKIITLDTATEIEGLSTGCKVYANIGEDTYDIVIPVKDDPDEGEARLEN
Aliases
michael-snyder:735
External resources

Characterizations

ZBTB33 (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody HPA005732. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 74.484
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ZBTB33 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2, using the antibody HPA005732. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 74.484
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ZBTB33 (Homo sapiens)
HEK293T
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T, using the antibody HPA005732. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 74.484
Reviewer comment
Band is not 50% intensity
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ZBTB33 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody HPA005732. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
ZBTB33 (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878, using the antibody HPA005732. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 74.484
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ZBTB33 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line K562 using the antibody HPA005732. 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: 74.484.
Submitter comment
See mass spec results
Reviewer comment
Multiple bands and band at expected size is ~50% of signal. Mass-spec identified ZBTB33 in band A
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ZBTB33 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
compliant
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using the antibody HPA005732, 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
CPSF6 binds to cleavage and polyadenylation RNA substrates and promotes RNA looping
Reviewer comment
Not detected in band B but not major immunoreactive band.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford