ENCAB000AMW

Antibody against Homo sapiens ZNF384, Mus musculus ZNF384

Homo sapiens
HeLa-S3, HEK293T, HepG2, K562
characterized to standards
Mus musculus
any cell type or tissue
partially characterized
Homo sapiens
any cell type or tissue
Status
released
Source (vendor)
Sigma
Product ID
HPA004051
Lot ID
A57874
Host
rabbit
Clonality
polyclonal
Purification
affinity
Antigen description
Immunogen: Zinc finger protein 384 recombinant protein epitope signature tag (PrEST)

Characterizations

ZNF384 (Homo sapiens)
HEK293T
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T, using the antibody HPA004051. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 63.0
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ZNF384 (Homo sapiens)
Method: immunoprecipitation
not reviewed
Caption
A. Western blots on nuclear lysates from cell lines GM12878 (Lane1), K562 (Lane2), HeLaS3 (Lane3), and HepG2 (Lane4). B. Immunoprecipitation was performed on nuclear lysates from HelaS3 cells using antibody HPA004051 against ZNF384. Lane1: Nuclear lysate. Lane 2: Unbound material from immunoprecipitation with HPA004051. Lane 3: Bound material from immunoprecipitation with HPA004051. Lane 4: Bound material from control immunoprecipitation with rabbit IgG. Arrow indicates band of expected size (~63 kD) that is efficiently enriched in the specifically immunoprecipitated fraction. Band indicated by * in HelaS3 immunoprecipitate is IgG light chains. A singke band of the correct weight band (~208 kD) is detected by Western blotting with HPA004051 in multiple human cell lines. Immunoprecipitation from HelaS3 nuclear lysate specifically enriches a single protein of ~63KD. Based on these observations, this antibody meets this ENCODE criterion.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
ZNF384 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
not reviewed
Caption
Immunoprecipitation of ZNF384 from K562 cells using HPA004051. Lane 1: input nuclear lysate, Lane 2: material immunoprecipitated with HPA004051, Lane 3: material immunoprecipitated using control IgG. Bands A was excised from the gel and subject to analysis by mass spectrometry. IP followed by mass spectrometry: Briefly, protein was immunoprecipitated from K562 whole cell lysates using NB100-68209, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGE Bis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was silver-stained, gel fragments corresponding to the bands indicated were excised and destained using the SilverSNAP Stain for Mass Spectrometry (Pierce). Then proteins were trypsinized using the in-gel digestion method. Digested proteins were analyzed on an LTQ-Orbitrap (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Protein false discovery rate < 1%, 2 peptides per protein minimum). We report 20 different proteins identified in band A, of this 9 were also detected in control IP indicating non specific enrichment during immunoprecipitation. Of the specifically immunoprecipitated proteins, ZNF384 is the most abundant protein in both band A . Based on these observations, this band is likely due to the presence of immunoprecipitated ZNF384 and HPA004051 meets the ENCODE standard for validation by this criterion.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
ZNF384 (Homo sapiens)
HeLa-S3
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
B. Immunoprecipitation was performed on nuclear lysates from HeLa-S3 cells using antibody HPA004051 against ZNF384. Lane1: Nuclear lysate. Lane 2: Unbound material from immunoprecipitation with HPA004051. Lane 3: Bound material from immunoprecipitation with HPA004051. Lane 4: Bound material from control immunoprecipitation with rabbit IgG. Arrow indicates band of expected size (~63 kD) that is efficiently enriched in the specifically immunoprecipitated fraction. Band indicated by * in HeLa-S3 immunoprecipitate is IgG light chains.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
ZNF384 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation of ZNF384 from K562 cells using HPA004051. Lane 1: input nuclear lysate, Lane 2: material immunoprecipitated with HPA004051, Lane 3: material immunoprecipitated using control IgG. Bands A was excised from the gel and subject to analysis by mass spectrometry.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
ZNF384 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
compliant
Caption
IP followed by mass spectrometry: Briefly, protein was immunoprecipitated from K562 whole cell lysates using NB100-68209, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGE Bis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was silver-stained, gel fragments corresponding to the bands indicated were excised and destained using the SilverSNAP Stain for Mass Spectrometry (Pierce). Then proteins were trypsinized using the in-gel digestion method. Digested proteins were analyzed on an LTQ-Orbitrap (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Protein false discovery rate < 1%, 2 peptides per protein minimum). We report 20 different proteins identified in band A, of this 9 were also detected in control IP indicating non specific enrichment during immunoprecipitation. Of the specifically immunoprecipitated proteins, ZNF384 is the most abundant protein in both band A . Based on these observations, this band is likely due to the presence of immunoprecipitated ZNF384 and HPA004051 meets the ENCODE standard for validation by this criterion.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
ZNF384 (Mus musculus)
Method: immunoprecipitation followed by mass spectrometry
not reviewed
Caption
HAP004051 is validated in K562 by IP-Mass-Spec. See validation documents submitted for Human cell lines for details.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
ZNF384 (Mus musculus)
Method: immunoprecipitation
not reviewed
Caption
Immunoprecipitation of CH12 and MEL nuclear extracts using anti-ZNF384 antibody (HPA004051) specifically and efficiently enriched a single band of the expected molecular weight of ZNF384 (~63 kD).
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
ZNF384 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2 using the antibody HPA004051. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 63.0.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
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