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Rad21

Manufactured by Fortis Life Sciences
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Rad21 is a laboratory equipment product designed for use in scientific research. It functions as a device for the detection and analysis of DNA damage and repair processes. The core function of Rad21 is to facilitate the study of cellular responses to various forms of DNA damage, such as those caused by radiation or chemical agents.

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Lab products found in correlation

Truseq chip sample preparation kit, by Illumina (2 mentions) Viia 7 system, by Thermo Fisher Scientific (2 mentions) Rnapii, by Cell Signaling Technology (2 mentions) Igg from rabbit serum, by Merck Group (2 mentions) Power sybr green kit, by Thermo Fisher Scientific (2 mentions) Spriselect beads, by Beckman Coulter (2 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (2 mentions) Nextseq 500 platform, by Illumina (2 mentions)

3 protocols using rad21

1

Fractionation and Western Blotting

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Nuclear and cytosolic fractions or total cell lysates were separated by SDS-PAGE. The proteins were transferred to nitrocellulose membranes and incubated with the primary antibodies RAD21 (BETHYL), Kap-1 (BETHYL) (nuclear fraction marker), and tubulin (SIGMA) (cytoplasmic fraction marker) as an internal control, and WAPL (Abcam). After removal of the unbound primary antibody, membranes were incubated with secondary antibody–peroxidase conjugate (Jackson), processed for detection by chemiluminescence (Advansta) and imaged on a ImageQuant LAS 4000. Protein levels were quantified and normalized to Kap-1 and tubulin using ImageJ software.
Perez S., Gevor M., Davidovich A., Kaspi A., Yamin K., Domovich T., Meirson T., Matityahu A., Brody Y., Stemmer S.M., El-Osta A., Haviv I., Onn I, & Gal-Tanamy M. (2019). Dysregulation of the cohesin subunit RAD21 by Hepatitis C virus mediates host–virus interactions. Nucleic Acids Research, 47(5), 2455-2471.
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2

ChIP-seq Protocol for Chromatin Binding

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Chromatin immunoprecipitation (ChIP) was performed as previously described (Letting et al., 2004 (link)). Antibodies include: CTCF (Millipore; 07-729), Rad21 (Bethyl Laboratories; A302-583A), mCherry (Abcam; Ab167453), RNAPII (Cell Signaling; Cat#14958), IgG from rabbit serum (Sigma; 15006). Quantitative polymerase chain reaction (qPCR) was performed using Power SYBR Green kit (Invitrogen; 4368577) with signals detected by ViiA7 System (Life Technologies). ChIP-seq libraries were prepared using Illumina’s TruSeq ChIP sample preparation kit (Illumina, Cat#IP-202-1012) according to manufacturer’s specifications, with the addition of size selection (left side at 0.9x, right side at 0.6x) using SPRIselect beads (Beckman Coulter, Cat#B23318). Library size was determined (average 351 bp, range 333-372 bp) using the Agilent Bioanalyzer 2100, followed by quantitation using real-time PCR using the KAPA Library Quant Kit for Illumina (KAPA Biosystems; Cat#KK4835). Libraries were then pooled and sequenced (1x75bp) on the Illumina NextSeq 500 platform according to manufacturer’s instructions. Bclfastq2 v 2.15.04 (default parameters) was used to convert reads to fastq.
Luan J., Xiang G., Gómez-García P.A., Tome J.M., Zhang Z., Vermunt M.W., Zhang H., Huang A., Keller C.A., Giardine B.M., Zhang Y., Lan Y., Lis J.T., Lakadamyali M., Hardison R.C, & Blobel G.A. (2021). Distinct properties and functions of CTCF revealed by a rapidly inducible degron system. Cell reports, 34(8), 108783.
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3

ChIP-seq Protocol for Chromatin Binding

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Chromatin immunoprecipitation (ChIP) was performed as previously described (Letting et al., 2004 (link)). Antibodies include: CTCF (Millipore; 07-729), Rad21 (Bethyl Laboratories; A302-583A), mCherry (Abcam; Ab167453), RNAPII (Cell Signaling; Cat#14958), IgG from rabbit serum (Sigma; 15006). Quantitative polymerase chain reaction (qPCR) was performed using Power SYBR Green kit (Invitrogen; 4368577) with signals detected by ViiA7 System (Life Technologies). ChIP-seq libraries were prepared using Illumina’s TruSeq ChIP sample preparation kit (Illumina, Cat#IP-202-1012) according to manufacturer’s specifications, with the addition of size selection (left side at 0.9x, right side at 0.6x) using SPRIselect beads (Beckman Coulter, Cat#B23318). Library size was determined (average 351 bp, range 333-372 bp) using the Agilent Bioanalyzer 2100, followed by quantitation using real-time PCR using the KAPA Library Quant Kit for Illumina (KAPA Biosystems; Cat#KK4835). Libraries were then pooled and sequenced (1x75bp) on the Illumina NextSeq 500 platform according to manufacturer’s instructions. Bclfastq2 v 2.15.04 (default parameters) was used to convert reads to fastq.
Luan J., Xiang G., Gómez-García P.A., Tome J.M., Zhang Z., Vermunt M.W., Zhang H., Huang A., Keller C.A., Giardine B.M., Zhang Y., Lan Y., Lis J.T., Lakadamyali M., Hardison R.C, & Blobel G.A. (2021). Distinct properties and functions of CTCF revealed by a rapidly inducible degron system. Cell reports, 34(8), 108783.
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