Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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 (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 (ab209101).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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 (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 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 (ab150077) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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 (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 (ab209101).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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 (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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

• IP

Unknown

Immunoprecipitation - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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) (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 (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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

All lanes:

Immunoprecipitation - Anti-Rad21 antibody [EPR22506-15] - ChIP Grade (<a href='/en-us/products/primary-antibodies/rad21-antibody-epr22506-15-chip-grade-ab217678'>ab217678</a>)

Predicted band size: 72 kDa

Observed band size: 70-135 kDa

false

• ChIP-seq

Lab

ChIP-sequencing - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

This data was developed using ab217678, the same antibody clone in a different buffer formulation.

Chromatin was prepared from MCF7 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab217678 [EPR22506-15]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

This data was developed using ab217678, the same antibody clone in a different buffer formulation.

Chromatin was prepared from MCF7 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab217678 [EPR22506-15]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP

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ChIP - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

• ChIP-seq

Lab

ChIP-sequencing - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

This data was developed using ab217678, the same antibody clone in a different buffer formulation.

Chromatin was prepared from MCF7 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab217678 [EPR22506-15]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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 (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 (ab209101).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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 (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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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 (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 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 (ab150077) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

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 (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 (ab209101).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217678).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

This data was developed using ab217678, the same antibody clone in a different buffer formulation.

Chromatin was prepared from 3T3-L1 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab217678 [EPR22506-15]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

This data was developed using ab217678, the same antibody clone in a different buffer formulation.

Chromatin was prepared from 3T3-L1 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab217678 [EPR22506-15]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

This data was developed using ab217678, the same antibody clone in a different buffer formulation.

Chromatin was prepared from 3T3-L1 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab217678 [EPR22506-15]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

This data was developed using the same antibody clone in a different buffer formulation (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 ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

This data was developed using the same antibody clone in a different buffer formulation (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 ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Rad21 antibody [EPR22506-15] - BSA and Azide free (AB254483)

This data was developed using the same antibody clone in a different buffer formulation (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 ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.