Rad21 Recombinant Rabbit Monoclonal Antibody [PSH19-52]
cat.: HA751716
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IP, IF-Cell
Clonality: Monoclonal
Clone number: PSH19-52
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 72 kDa
Isotype: IgG
Positive control: HeLa cell lysate, MCF7 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, human breast cancer tissue, human colon tissue, mouse colon tissue, rat colon tissue, HeLa, NIH/3T3, C6.
Subcellular location: Nucleus, Nucleus matrix, Chromosome, centromere, Cytoplasm, cytoskeleton, spindle pole; Cytoplasm, cytosol, Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  IP
  IF-Cell
1:5,000
1:500-1:2,000
1-2μg/sample
1:500
Uniprot #: SwissProt: O60216 Human | Q61550 Mouse
Entrez Gene: 314949 Rat
Alternative names: CDLS4 Double-strand-break repair protein rad21 homolog hHR21 HR21 HRAD21 KIAA0078 MCD1 Nuclear matrix protein 1 NXP-1 NXP1 Protein involved in DNA double-strand break repair RAD21 RAD21 homolog (S. pombe) RAD21 homolog RAD21_HUMAN Scc1 SCC1 homolog
Images
HA751716_1.jpg Fig1: Western blot analysis of Rad21 on different lysates with Rabbit anti-Rad21 antibody (HA751716) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: RAW264.7 cell lysate
Lane 5: C6 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 72 kDa
Observed band size: 120 kDa

Exposure time: 11 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751716) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA751716_2.jpg Fig2: Western blot analysis of Rad21 on different lysates with Rabbit anti-Rad21 antibody (HA751716) at 1/5,000 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 72 kDa
Observed band size: 120 kDa

Exposure time: 1 minute 18 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751716) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA751716_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Rad21 antibody (HA751716) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751716) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA751716_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Rad21 antibody (HA751716) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751716) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA751716_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Rad21 antibody (HA751716) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751716) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA751716_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-Rad21 antibody (HA751716) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751716) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA751716_7.jpg Fig7: Rad21 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA751716 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751716 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA751716 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA751716 in HeLa cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 10 seconds; ECL: K1801
HA751716_8.jpg Fig8: Immunocytochemistry analysis of HeLa cells labeling Rad21 with Rabbit anti-Rad21 antibody (HA751716) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Rad21 antibody (HA751716) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA751716_9.jpg Fig9: Immunocytochemistry analysis of NIH/3T3 cells labeling Rad21 with Rabbit anti-Rad21 antibody (HA751716) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Rad21 antibody (HA751716) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA751716_10.jpg Fig10: Immunocytochemistry analysis of C6 cells labeling Rad21 with Rabbit anti-Rad21 antibody (HA751716) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Rad21 antibody (HA751716) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.