Recombinant Anti-Rad51 antibody [EPR4030(3)] - BSA and Azide free (ab221796)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4030(3)] to Rad51 - BSA and Azide free
- Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
- Reacts with: Mouse, Rat, Human
Overview
-
Product name
Anti-Rad51 antibody [EPR4030(3)] - BSA and Azide free
See all Rad51 primary antibodies -
Description
Rabbit monoclonal [EPR4030(3)] to Rad51 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Rad51.
(Peptide available asab176353) -
Positive control
- WB: C6, NIH/3T3, 293T, Jurkat, HeLa, and K562 cell lysates and mouse spleen tissue lysate. IHC-P: Human cervix carcinoma, lung and testis tissues. ICC/IF: Jurkat cells. Flow Cyt: HeLa cells. IP: HEK293 cell lysates.
-
General notes
Ab221796 is the carrier-free version of ab133534. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
ab221796 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Dissociation constant (KD)
KD = 9.20 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4030(3) -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Positive Controls
- HeLa whole cell lysate (ab150035)
- Mouse spleen tissue lysate - total protein (ab29293)
- HeLa whole cell lysate (ab29545)
- Jurkat whole cell lysate (ab30128)
- NIH 3T3 whole cell lysate (ab7179)
- Mouse spleen tissue lysate - total protein (14 days) (ab7274)
- Mouse spleen tissue lysate - total protein (7 days) (ab7275)
- Jurkat whole cell lysate (ab7899)
- HEK293 whole cell lysate (ab7902)
- K562 whole cell lysate (ab7911)
- Mouse small intestine tissue lysate - total protein (ab7939)
-
Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab221796 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
WB | Use at an assay dependent concentration. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa). | |
IP | Use at an assay dependent concentration. | |
IHC-P | Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
Flow Cyt | Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
ICC/IF | Use at an assay dependent concentration. |
Target
-
Function
Plays an important role in homologous strand exchange, a key step in DNA repair through homologous recombination. Binds to single and double-stranded DNA and exhibits DNA-dependent ATPase activity. Catalyzes the recognition of homology and strand exchange between homologous DNA partners to form a joint molecule between a processed DNA break and the repair template. Binds to single-stranded DNA in an ATP-dependent manner to form nucleoprotein filaments which are essential for the homology search and strand exchange (PubMed:26681308). Part of a PALB2-scaffolded HR complex containing BRCA2 and RAD51C and which is thought to play a role in DNA repair by HR. Plays a role in regulating mitochondrial DNA copy number under conditions of oxidative stress in the presence of RAD51C and XRCC3. -
Tissue specificity
Highly expressed in testis and thymus, followed by small intestine, placenta, colon, pancreas and ovary. Weakly expressed in breast. -
Involvement in disease
Breast cancer
Mirror movements 2
Defects in RAD51 are found in a patient with microcephaly, mental retardation without bone marrow failure and pediatric cancers. -
Sequence similarities
Belongs to the RecA family. RAD51 subfamily.
Contains 1 HhH domain. -
Domain
The nuclear localization may reside in the C-terminus (between 259 and 339 AA). -
Post-translational
modificationsUbiquitinated by the SCF(FBXO18) E3 ubiquitin ligase complex, regulating RAD51 subcellular location and preventing its association with DNA.
Phosphorylated. Phosphorylation of Thr-309 by CHEK1 may enhance association with chromatin at sites of DNA damage and promote DNA repair by homologous recombination. Phosphorylation by ABL1 inhibits function. -
Cellular localization
Nucleus. Cytoplasm. Cytoplasm, perinuclear region. Mitochondrion matrix. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. Colocalizes with RAD51AP1 and RPA2 to multiple nuclear foci upon induction of DNA damage. DNA damage induces an increase in nuclear levels. Together with FIGNL1, redistributed in discrete nuclear DNA damage-induced foci after ionizing radiation (IR) or camptothecin (CPT) treatment. Accumulated at sites of DNA damage in a SPIDR-dependent manner. - Information by UniProt
-
Database links
- Entrez Gene: 5888 Human
- Entrez Gene: 19361 Mouse
- Entrez Gene: 499870 Rat
- Omim: 179617 Human
- SwissProt: Q06609 Human
- SwissProt: Q08297 Mouse
- Unigene: 631709 Human
- Unigene: 330492 Mouse
see all -
Alternative names
- BRCA1/BRCA2 containing complex, subunit 5 antibody
- BRCC 5 antibody
- BRCC5 antibody
see all
Images
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [EPR4030(3)] - BSA and Azide free (ab221796)
Immunohistochemistry (PFA-fixed paraffin-embedded sections) analysis of human colonic carcinoma tissue labelling with ab221796 at 0.54µg/mL. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer pH 9). A HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody. Counterstained with hematoxylin.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [EPR4030(3)] - BSA and Azide free (ab221796)
Immunohistochemistry (PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling with ab221796 at 0.54µg/mL. Heat mediated antigen retrieval was performed using ab92684 (Tris/EDTA buffer pH 9). A HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody. Counterstained with hematoxylin.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [EPR4030(3)] - BSA and Azide free (ab221796)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling Rad51 with purified ab133534 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133534).
-
Immunocytochemistry/ Immunofluorescence - Anti-Rad51 antibody [EPR4030(3)] - BSA and Azide free (ab221796)
Immunocytochemistry/Immunofluorescence analysis of Jurkat (human T cell leukemia cell line from peripheral blood) cells labelling Rad51 with purified ab133534 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/1000) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133534).
-
Flow Cytometry analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling Rad51 with purified ab133534 at 1/350 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133534).
-
ab133534 (purified) at 1/100 immunoprecipitating Rad51 in HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate.
Lane 1 (input): HEK-293 whole cell lysate (10µg)
Lane 2 (+): ab133534 + HEK-293 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133534 in HEK-293 whole cell lysate.
For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133534).
-
Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with unpurified ab133534 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab133534, 1/1000 dilution) for 30 minutes at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1 μg/1x106 cells) used under the same conditions. Unlabeled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133534).
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [EPR4030(3)] - BSA and Azide free (ab221796)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling Rad51 with unpurified ab133534 at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133534).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133534).
Protocols
Datasheets and documents
Certificate of Compliance
References (2)
ab221796 has been referenced in 2 publications.
- Boysen G et al. SPOP mutation leads to genomic instability in prostate cancer. Elife 4:N/A (2015). WB . PubMed: 26374986
- Shen Y et al. BMN 673, a novel and highly potent PARP1/2 inhibitor for the treatment of human cancers with DNA repair deficiency. Clin Cancer Res 19:5003-15 (2013). ICC/IF . PubMed: 23881923