Anti-Rad21 antibody - ChIP Grade [EPR22506-15] (ab217678)

Rabbit Recombinant Monoclonal RAD21 antibody. Suitable for IP, ChIP, WB, ICC/IF, Flow Cyt (Intra), IHC-P, ChIC/CUT&RUN-seq and reacts with Human, Mouse, Rat samples. Cited in 2 publications.

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Images

ChIP - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (AB217678), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	ChIP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P	ChIC/CUT&RUN-seq
Human	Tested	Tested	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Expected	Tested	Tested	Tested	Tested	Expected
Rat	Expected	Expected	Tested	Expected	Expected	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 5 µg	Notes 5 μg Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes -
Species Human	Dilution info 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000	Notes Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 5 µg	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information RAD21

Function

Double-strand-break repair protein rad21 homolog. As a member of the cohesin complex, involved in sister chromatid cohesion from the time of DNA replication in S phase to their segregation in mitosis, a function that is essential for proper chromosome segregation, post-replicative DNA repair, and the prevention of inappropriate recombination between repetitive regions (PubMed:11509732). The cohesin complex may also play a role in spindle pole assembly during mitosis (PubMed:11590136). In interphase, cohesins may function in the control of gene expression by binding to numerous sites within the genome (By similarity). May control RUNX1 gene expression (Probable). Binds to and represses APOB gene promoter (PubMed:25575569). May play a role in embryonic gut development, possibly through the regulation of enteric neuron development (By similarity). 64-kDa C-terminal product. May promote apoptosis.

Alternative names

HR21, KIAA0078, NXP1, SCC1, RAD21, Double-strand-break repair protein rad21 homolog, hHR21, Nuclear matrix protein 1, SCC1 homolog, NXP-1

Recommended products

Rabbit Recombinant Monoclonal RAD21 antibody. Suitable for IP, ChIP, WB, ICC/IF, Flow Cyt (Intra), IHC-P, ChIC/CUT&RUN-seq and reacts with Human, Mouse, Rat samples. Cited in 2 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR22506-15
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Rad21 also known as double-strand-break repair protein rad21 homolog serves as a component of the cohesin complex. This protein is approximately 123 kDa in size. Rad21 is expressed in a wide range of tissues and cell types particularly in those with high proliferative activity. The protein plays an essential mechanical role by facilitating the cohesion of sister chromatids during cell division ensuring successful chromosome segregation.

Biological function summary

Rad21 is essential in processes beyond chromatid cohesion. It forms part of the cohesin complex which also includes proteins like SMC1 SMC3 and SCC3/SA1 or SA2. This complex is central to maintaining genome stability by participating in DNA repair and regulating gene expression. Through these roles Rad21 contributes to cellular processes such as cell cycle progression and embryonic development.

Pathways

Rad21 integrates into the DNA damage response and cell cycle regulation pathways. The cohesin complex with Rad21 as an important factor acts alongside other proteins such as ATM and ATR in the DNA damage response ensuring the repair of double-strand breaks. Within the cell cycle Rad21 helps coordinate the timing and accuracy of cell division working with cyclin-dependent kinases.

Associated diseases and disorders

Rad21 has been linked to cancer and Cornelia de Lange syndrome. Mutations or dysregulation of Rad21 can result in tumorigenesis due to errors in chromosome segregation and compromised DNA repair. In Cornelia de Lange syndrome cohesin-related proteins like NIPBL and SMC1A also play a role indicating a disruption in the cohesin pathway's function. Understanding Rad21's involvement helps in developing targeted therapies for these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

16 product images

ChIP - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Chromatin was prepared from MCF7 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min.
The ChIP was performed with 25 μg of chromatin, 5 μg of ab217678 (red), and 20 μl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
The observed enrichment profile of Rad21 binding is consistent with what has been described in the literature (PMID: 23145106)
Western blot - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Full length Rad21 is approximately 130kDa, cleaved Rad21 is approximately 75 kDa. The molecular weight observed are consistent with what has been described in the literature (PMID: 12417729, PMID 21876002).
Exposure time: 7.7 seconds
Blocking/diluting buffer and concentration: 5% NFDM/TBST
All lanes: Western blot - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678) at 1/1000 dilution
Lane 1: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 3: C6 (rat glial tumor glial cell), whole cell lysate whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 72 kDa
Observed band size: 130-75 kDa
Western blot - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Full length Rad21 is approximately 130kDa, cleaved Rad21 is approximately 75 kDa. The molecular weight observed are consistent with what has been described in the literature (PMID: 12417729, PMID 21876002).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes.
All lanes: Western blot - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678) at 1/1000 dilution
Lane 1: Rat Brain tissue lysate at 20 µg
Lane 2: Mouse Brain tissue lysate at 20 µg
Lane 3: Mouse heart tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 72 kDa
Observed band size: 130-75 kDa
Western blot - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Full length Rad21 is approximately 130 kDa, cleaved Rad21 is approximately 75 kDa. The molecular weight observed are consistent with what has been described in the literature (PMID: 12417729, PMID 21876002).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Exposure times:
Lanes1-2: 37 seconds
Lane 3: 5.5 seconds
Lane 4: 48 seconds
All lanes: Western blot - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678) at 1/1000 dilution
Lane 1: Human heart tissue lysate at 20 µg
Lane 2: Human fetal kidney tissue lysate at 20 µg
Lane 3: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 4: MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
Lanes 1 - 2: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution
Lanes 3 - 4: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 72 kDa
Observed band size: 130-75 kDa
Immunoprecipitation - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Rad21 was immunoprecipitated from 0.35mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab217678 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab217678 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used as the secondary antibody at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10μg (input)
Lane 2: ab217678 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab217678 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
Faint band below 75kDa in lane 2 could be Rad21 cleavage product as described in the literature (PMID: 28854249).
All lanes: Immunoprecipitation - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Predicted band size: 72 kDa
Observed band size: 70-135 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Rad21 using ab217678 at 1/1000 dilution for 30 minutes at room temperature, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on rat colon (PMID: 25575569) is observed. Counterstained with Hematoxylin. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Rad21 using ab217678 at 1/1000 dilution for 30 minutes at room temperature, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on mouse stomach (PMID: 25575569) is observed. Counterstained with Hematoxylin. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling Rad21 using ab217678 at 1/1000 dilution for 30 minutes at room temperature, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on human stomach (PMID: 25575569) is observed. Counterstained with Hematoxylin. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Rad21 using ab217678 at 1/1000 dilution for 30 minutes at room temperature, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on the human breast carcinoma (PMID: 21255398) is observed. Counterstained with Hematoxylin. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling Rad21 using ab217678 at 1/500 dilution (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730, Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling Rad21 with ab217678 at 1/100 dilution, followed by an AlexaFluor^®488 Goat anti-Rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution (green). Confocal image showing nuclear staining in NIH/3T3 cell line is observed. The nuvlear counterstain is DAPI (Blue). Tubulin is counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is an AlexaFluor^®488 Goat anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Rad21 with ab217678 at 1/100 dilution, followed by an AlexaFluor^®488 Goat anti-Rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution (green). Confocal image showing nuclear staining in HeLa cell line is observed. The nuvlear counterstain is DAPI (Blue). Tubulin is counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is an AlexaFluor^®488 Goat anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
ChIC/CUT&RUN sequencing - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human epithelial cells from cervix adenocarcinoma) cells and 5 µg of ab217678 [EPR22506-15]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
ChIC/CUT&RUN sequencing - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human epithelial cells from cervix adenocarcinoma) cells and 5 µg of ab217678 [EPR22506-15]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
ChIC/CUT&RUN sequencing - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human epithelial cells from cervix adenocarcinoma) cells and 5 µg of ab217678 [EPR22506-15]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
Flow Cytometry (Intracellular) - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (ab217678)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (mouse embryonic fibroblast) cell line labeling Rad21 using ab217678 at 1/500 dilution (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730, Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.