ENCAB292USO
Antibody against Homo sapiens ZBTB33
Homo sapiens
K562, GM12878, MCF-7, HepG2
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
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
- Grant
- U54HG006996
- Download
- 1109_07_ZBTB33_HPA005732.jpg
ZBTB33 (Homo sapiens)
HepG2
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
- Grant
- U54HG006996
- Download
- 1109_08_ZBTB33_HPA005732.jpg
ZBTB33 (Homo sapiens)
HEK293T
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
- Grant
- U54HG006996
- Download
- 1112_08_ZBTB33_HPA005732.jpg
ZBTB33 (Homo sapiens)
K562
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
- Grant
- U54HG006996
- Download
- Exp.805_1-ZBTB33.jpg
ZBTB33 (Homo sapiens)
GM12878
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
- Grant
- U54HG006996
- Download
- expt1110_6-ZBTB33-HPA005732.jpg
ZBTB33 (Homo sapiens)
K562
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
- Grant
- U54HG006996
- Download
- ZBTB33 (HPA005732).JPG
ZBTB33 (Homo sapiens)
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
- Grant
- U54HG006996
- Download
- ZBTB33_HPA005732 final.pdf