1. Antibodies
  2. Homo sapiens
  3. RAD21

ENCAB000BSF

Antibody against Homo sapiens RAD21

Homo sapiens
K562, GM12878, GM12892, GM19238, GM19240, GM12891, GM19239
characterized to standards with exemption
Status
released
Source (vendor)
Abcam
Product ID
ab992
Lot ID
GR106791
Characterized targets
RAD21 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Isotype
IgG
Antigen description
Synthetic peptide (Human) conjugated to KLH - which represents a portion of human Rad21 encoded within exon 14
External resources

Characterizations

RAD21 (Homo sapiens)
Method: knockdown or knockout
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
A new secondary characterization is not required as one exists for ENCAB000AKG which is a lot of the same antibody (ab992). The primary characterization (https://www.encodeproject.org/antibody-characterizations/2bf7eeee-2a31-46ba-956d-ecac94d65994/) of K562 for this lot shows the same banding pattern as the primary characterization (https://www.encodeproject.org/antibody-characterizations/a998bebe-57f7-4de9-8732-735fd07a8764) in K562 cell line for the lot ENCAB000AKG.
Submitter comment
This is a new lot of a previously characterized antibody ENCAB000AKG and we think no further characterization is necessary beyond the primary if the banding pattern is consistent with the old.
Reviewer comment
The banding pattern in GM12878 and K562 is consistent with that of the primary characterizations associated with old lot ENCAB000AKG so a new secondary characterization for the new lot is not needed.
Submitted by
Jessika Adrian
Lab
Michael Snyder, Stanford
RAD21 (Homo sapiens)
GM12878GM12892GM19238GM19240GM12891GM19239
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878, using the antibody ab992. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG. Molecular Weight: 71.69, band observed at 130KD
Submitter comment
This is the same banding pattern found in ENCAB529YRC and ENCAB770AFW and ENCAB000AKG. While the predicted size of Rad21 is ~72kD, according to product characterization literature (found at manufacturer'€™s website), the bands at ~70kD are of unknown origin while a band of ~140kD, similar to the one observed here, is identified as Rad21. In support of this, siRNAs targeting Rad21 dramatically reduce levels of the 140kD band, while levels of the 70kD band are unaffected (see Figure 2 of this document). 1. Hyperphosphorylation (ref. PMID 11073952 & many others) https://www.ncbi.nlm.nih.gov/gene/5885 2. Dimerisation as part of cohesin complex (whose other members we can detect in the 130kDa band, refs. PMIDs 23874961 & 19075111)
Reviewer comment
Multiple bands and band at expected size not >50% of total signal in lane. Peggy Farham has said that one GM is equivalent to all GM cell lines so that if you characterized in one line, all lines are considered characterized.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
RAD21 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody ab992. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG. Molecular Weight: 71.69, band observed at 130KD
Submitter comment
The banding pattern here is the same as found in ENCAB770AFW and ENCAB000AKG and ENCAB529YRC. While the predicted size of Rad21 is ~72kD, according to product characterization literature (found at manufacturer'€™s website), the bands at ~70kD are of unknown origin while a band of ~140kD, similar to the one observed here, is identified as Rad21. In support of this, siRNAs targeting Rad21 dramatically reduce levels of the 140kD band, while levels of the 70kD band are unaffected (see Figure 2 of this document). 1. Hyperphosphorylation (ref. PMID 11073952 & many others) https://www.ncbi.nlm.nih.gov/gene/5885 2. Dimerisation as part of cohesin complex (whose other members we can detect in the 130kDa band, refs. PMIDs 23874961 & 19075111)
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
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 K562-Rad21.jpg