1. Antibodies
  2. Homo sapiens
  3. HDGF

ENCAB691BTB

Antibody against Homo sapiens HDGF

Homo sapiens
K562
characterized to standards
Status
released
Source (vendor)
Bethyl Labs
Product ID
A303-169A
Lot ID
1
Characterized targets
HDGF (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1026
External resources

Characterizations

HDGF (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody A303-169A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Submitter comment
HDGF is known to form a domain-swapped homodimer that can be stable even in reducing gels (PMID: 17270212)
Reviewer comment
Higher than expected size and multiple bands were noted. In response to the submitter's comment, the stable domain-swapped homodimer would explain the higher (double-sized) band and also explain why the ratio of the two bands can be different on different westerns.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
HDGF (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation of HDGF from K562 cells using A303-169A. Lane 1: input nuclear lysate. Lane 2: material immunoprecipitated with A303-169A. Lane 3: material immunoprecipitated using control IgG. Band A was excised from gel and subject to analysis by mass spectrometry. The expected band size is 28 kDa.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
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 HDGF.jpeg
HDGF (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
IP followed by mass spectrometry: Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using A303-169A, 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
Kathrina Onate
Lab
Michael Snyder, Stanford