ENCAB014HCW

Antibody against Homo sapiens PPP1R10

Homo sapiens
GM12878, HEK293T
characterized to standards
Homo sapiens
K562
characterized to standards with exemption
Homo sapiens
HepG2, MCF-7
not characterized to standards
Status
released
Source (vendor)
Bethyl Labs
Product ID
A300-439A
Lot ID
2
Characterized targets
PPP1R10 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1558
External resources

Characterizations

PPP1R10 (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody A300-439A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 99.058
Reviewer comment
Multiple bright bands
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
PPP1R10 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2, using the antibody A300-439A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 99.058
Reviewer comment
Band of interest is not 50% of overall signal in lane, and multiple bands appear in both input and the lane of interest.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
PPP1R10 (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 A300-439A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 99.058
Submitter comment
We analyzed the bands by IP-MS and confirmed that the factor of interest is detected.
Reviewer comment
There are multiple immunoreactive bands.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
PPP1R10 (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878, using the antibody A300-439A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 99.058
Submitter comment
We did 4 lanes as usual and the lane between IP and IgG is because the film shifted or IP lane was too full so 1 or 2 ul flew to the next lane.
Reviewer comment
Not sure about extra band on lane 4
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
PPP1R10 (Homo sapiens)
HEK293T
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T, using the antibody A300-439A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 99.058
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
PPP1R10 (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 A300-439A, 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).
Submitted by
Jessika Adrian
Lab
Michael Snyder, Stanford
PPP1R10 (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 A300-439A. 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: 99kDa
Submitter comment
We confirmed the multiple bands seen in the IP via mass spec.
Reviewer comment
The TF of interest was detected in marked bands A, B and C by mass spectrometry.
Submitted by
Jessika Adrian
Lab
Michael Snyder, Stanford