Name: Anti-Rad51 antibody [14B4]
SKU: ab213
Rating: 4 (9 reviews)

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Western blot - Anti-Rad51 antibody [14B4] (ab213)

Key features and details

Mouse monoclonal [14B4] to Rad51
Suitable for: IHC-P, WB
Reacts with: Human
Isotype: IgG2b

Product name

Anti-Rad51 antibody [14B4]
See all Rad51 primary antibodies
Description

Mouse monoclonal [14B4] to Rad51
Host species

Mouse
Specificity

Specific for human Rad51.

Tested Applications & Species

Application	Species
IHC-P	Human
WB	Human

See all applications and species data

Immunogen

Recombinant full length protein corresponding to Human Rad51 aa 1-338.
Database link: Q06609
Positive control
- WB: RAD51 shRNA transfected (+) 293T whole cell extracts, Jurkat, Raji and NCI-H929 whole cell lysate. IHC-P: BT483 xenoograft tissue, human colon esophagus tissue.
General notes

This product was changed from ascites to tissue culture supernatant on 17^th September 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer

pH: 7.40
Constituent: 100% PBS
Concentration information loading...
Purity

Immunogen affinity purified
Clonality

Monoclonal
Clone number

14B4
Myeloma

NS1
Isotype

IgG2b
Light chain type

kappa
Research areas

Compatible Secondaries
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
- Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Isotype control
- Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
Recombinant Protein
- Recombinant human Rad51 protein (Active) (ab63808)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab213 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
IHC-P	Human
WB	Human

Application	Abreviews	Notes
IHC-P	(1)	1/100 - 1/1000.
WB	(5)	1/500 - 1/3000. Predicted molecular weight: 37 kDa. Please see WB protocol in image legend.

Notes
IHC-P 1/100 - 1/1000.
WB 1/500 - 1/3000. Predicted molecular weight: 37 kDa. Please see WB protocol in image legend.

Application notes

Is unsuitable for ICC/IF.

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
modifications

Ubiquitinated 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.
Target information above from: UniProt accession Q06609 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 5888 Human
- Omim: 179617 Human
- SwissProt: Q06609 Human
- Unigene: 631709 Human
Alternative names
- BRCA1/BRCA2 containing complex, subunit 5 antibody
- BRCC 5 antibody
- BRCC5 antibody
- DNA repair protein RAD51 homolog 1 antibody
- DNA repair protein rhp51 antibody
- FANCR antibody
- hRAD51 antibody
- HsRAD51 antibody
- HsT16930 antibody
- MRMV2 antibody
- Rad 51 antibody
- RAD51 antibody
- RAD51 homolog (RecA homolog, E. coli) (S. cerevisiae) antibody
- RAD51 homolog A antibody
- RAD51 homolog antibody
- RAD51 recombinase antibody
- RAD51, S. cerevisiae, homolog of antibody
- RAD51_HUMAN antibody
- RAD51A antibody
- RECA antibody
- RecA like protein antibody
- RecA, E. coli, homolog of antibody
- Recombination protein A antibody
see all

Western blot - Anti-Rad51 antibody [14B4] (ab213)

All lanes : Anti-Rad51 antibody [14B4] (ab213) at 1/500 dilution

Lane 1 : Jurkat whole cell lysate
Lane 2 : Raji whole cell lysate
Lane 3 : NCI-H929 whole cell lysate

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-mouse IgG antibody at 1/10000 dilution

Developed using the ECL technique.

Predicted band size: 37 kDa

10% SDS-PAGE

Running condition: 80V, 15min; 140V, 40 min

Transfer condition: Semi-dry, 18 V, 60 min (Nitrocellulose membrane)

Blocking condition: 5% non-fat milk in TBST, RT, 60 min.

Primary antibody incubation: 4°C overnight.

Washing condition: 5 ml TBST, 4 x 5 min.

Chemiluminescent HRP Substrate.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Rad51 with ab213 at 1/100 dilution at 4°C overnight. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Endogenous peroxidase blocking: 3% H₂O₂, RT, 30minutes. Blocking: 1.5% goat serum (dilute goat serum by 1xPBS), RT, 30 min. Secondary antibody: ABC HRP Kit (Mouse IgG), 1:200, RT, 30min. Washing: PBS, 2 x 5 mins. Chromogen system: DAB.
Western blot - Anti-Rad51 antibody [14B4] (ab213)

All lanes : Anti-Rad51 antibody [14B4] (ab213) at 1/500 dilution

Lane 1 : Non-transfected 293T (-) whole cell extracts
Lane 2 : RAD51 shRNA transfected (+) 293T whole cell extracts

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-mouse IgG antibody

Developed using the ECL technique.

Predicted band size: 37 kDa

10% SDS-PAGE
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)

Immunohistochemical analysis of paraffin-embedded human esophagus tissue labeling Rad51 with ab213 at 1/100 dilution at 4°C overnight. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Endogenous peroxidase blocking: 3% H₂O₂, RT, 30minutes. Blocking: 1.5% goat serum (dilute goat serum by 1xPBS), RT, 30 min. Secondary antibody: ABC HRP Kit (Mouse IgG), 1:200, RT, 30min. Washing: PBS, 2 x 5 mins. Chromogen system: DAB.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)

Immunohistochemical analysis of paraffin-embedded BT483 xenoograft tissue labeling RAD51 with ab213 at 1/200 dilution. Nuclear staining is observed. Antigen Retrieval: EDTA based, pH 8.0 buffer, 15min.
Western blot - Anti-Rad51 antibody [14B4] (ab213)Image courtesy of an anonymous Abreview.

Anti-Rad51 antibody [14B4] (ab213) at 1/500 dilution + lysate prepared from murine NIH3T3 cells at 20 µg

Secondary
HRP conjugated anti-mouse IgG at 1/5000 dilution

Developed using the ECL technique.

Predicted band size: 37 kDa
Observed band size: 42,45 kDa
why is the actual band size different from the predicted?

Exposure time: 5 minutes
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)This image is courtesy of an anonymous Abreview

ab213 staining Rad51 in Human Lovo xenograft tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 20% serum for 20 minutes at room temperature; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/100 in TBST) for 1 hour. An undiluted HRP-conjugated Donkey anti-mouse IgG polyclonal was used as the secondary antibody.
See Abreview

Western blot protocols
Immunohistochemistry protocols

Click here to view the general protocols

Publishing research using ab213? Please let us know so that we can cite the reference in this datasheet.

ab213 has been referenced in 69 publications.

Salmina K et al. "Mitotic Slippage" and Extranuclear DNA in Cancer Chemoresistance: A Focus on Telomeres. Int J Mol Sci 21:N/A (2020). PubMed: 32316332
Horikoshi N et al. Pre-existing H4K16ac levels in euchromatin drive DNA repair by homologous recombination in S-phase. Commun Biol 2:253 (2019). PubMed: 31286070
Silva B et al. FANCM limits ALT activity by restricting telomeric replication stress induced by deregulated BLM and R-loops. Nat Commun 10:2253 (2019). PubMed: 31138795
Sahin ID et al. Crizotinib and PARP inhibitors act synergistically by triggering apoptosis in high-grade serous ovarian cancer. Oncotarget 10:6981-6996 (2019). PubMed: 31857852
Huang TH et al. The Histone Chaperones ASF1 and CAF-1 Promote MMS22L-TONSL-Mediated Rad51 Loading onto ssDNA during Homologous Recombination in Human Cells. Mol Cell 69:879-892.e5 (2018). PubMed: 29478807

View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A

Submit a review Submit a question

1-10 of 20 Abreviews or Q&A

Application

Western blot

Loading amount

20 µg

Gel Running Conditions

Non-reduced Denaturing (15%)

Sample

Human Cell lysate - whole cell (293T)

Specification

293T

Blocking step

Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jan 15 2015

Application

Immunocytochemistry/ Immunofluorescence

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C

Sample

Human Cell (293T)

Specification

293T

Permeabilization

Yes - 0.5% Triton X100

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 13 2015

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Blocking step

Serum as blocking agent for 20 minute(s) · Concentration: 20% · Temperature: RT°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: pH9

Sample

Human Tissue sections (Lovo Xenograft)

Specification

Lovo Xenograft

Permeabilization

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Jul 17 2013

Question

The customer has responded as below;

Please could we have a credit note.

In regards to the ab58101 antibody, I think it is generally accepted that full length Sash1 runs at 170 kDa, however another band can be observed at 130kDa in some samples (see attached paper). However, using ab58101 neither the 170 kDa or 130 kDa band can be detected, whereas the Sash1 antibody from one of your competitors detects both bands in the same sample.

Thanks for your help and hope to hear from you soon

Kind regards,

Abcam community

Verified customer

Asked on Nov 29 2012

Answer

Thank you for contacting us.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Abcam Scientific Support

Answered on Nov 29 2012

Question

Please see below for the response from the customer

I have attached western images for the SETBP1 and Sash1 antibodies. Unfortunately I didn't save any of the IF images for the Rad51 antibody as the quality was too bad.

Thanks!

Kind regards,

Abcam community

Verified customer

Asked on Nov 28 2012

Answer

Thank you for your email.

I would be happy to process refunds for ab213 and ab68715. Could you however let me know why the customer expecting a 170kDa band for ab58101 which should be nearly 130-140 kDa.

Thanks!

Abcam Scientific Support

Answered on Nov 28 2012

Question

Please see below for an email we have received from a customer;

I've ordered several antibodies from AbCam via Sapphire in the last few months and on the whole the quality has been very good. However, we have had three antibodies that have not worked at all for western blot or immunofluorescence and were wondering whether we could get a refund on these three?

Ab213 Rad51 antibody – this antibody shows a lot of background and cytoplasmic staining on immunofluorescence at the recommended concentration and lower and we are unable to see DNA repair foci following DNA damage induction via IR.

Sapphire Order #: PO-15316
Abcam Invoice #: 1348460E

AB68715 SETBP1 antibody – we are unable to detect a band of the expected size of around 170 kDa on western blot.

Sapphire Order #: PO-15671

Abcam Invoice #: 1353502-SAP002E

AB58101 SASH1 antibody – we were unable to detect a band at the correct size of around 170 kDa via western blot or any staining via IF.

Sapphire Order #: PO-15316

Abcam Invoice #: 1347756F

All of these antibodies have been trialled by at least two experienced people in our lab and western blotting and immunofluorescence are techniques we have extensive experience with. I can provide further details of the protocols used if required.

Thanks for your help!

Kind regards,

Abcam community

Verified customer

Asked on Nov 21 2012

Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

- Could you provide us the images? we may help you with the troubleshooting.

Thank you very much for your cooperation. I will look forward to hearing from you soon.

Abcam Scientific Support

Answered on Nov 21 2012

Question

I need your help to address the problem, the customer us Abcam( cat #: ab213) primary antibody for western blotting, and the experiment didn’t work, could you help him find out what is the problem? Please see the attached for the protocol he follows ,

Thanks a lot , your quick response will be greatly appreciated!

Abcam community

Verified customer

Asked on May 17 2012

Answer

Thank you for contacting Abcam.

I just had a few more protocol questions for you regarding the problems you are having with ab213 in western blot.

1 - Why was 2 different lysis buffers used in the extraction protocol? Either of them should have been suffcient on their own.

2 - Whenyou say that the wetern blotting did not work. Does that means there was no signal, high background, weak signal?

3 - Could you please provide the lot number of the antibody you are using?

I look forward to your reply and helping resolve this situation.

Abcam Scientific Support

Answered on May 17 2012

Question

Thank you so much for your help. I will sent you update of the ab63801. And the secondary antibody we used was 1:2000 for ab213.

Abcam community

Verified customer

Asked on Feb 27 2012

Answer

I am happy to help. Best of luck in your experiments!

Abcam Scientific Support

Answered on Feb 27 2012

Question

I have used this antibody to stain MCF7 cells successfully via ICC/IF but WB with MCF7 lysate from 50ug protein extract, with primary at 1:250 and 1:500, barely showed a band after 45' exposure

Abcam community

Verified customer

Asked on Feb 27 2012

Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab63801.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Abcam Scientific Support

Answered on Feb 27 2012

Question

ICC: sees big red blob in nucleus (correct location) MCF7 cells, treatment with mitomicine C Ab:1/500 1h secondary: 1/200 Texas red (goat anti-mouse), works well otherwise pre-extraction with 0.25% TX fixation: 3% PFA perm step: 0.5% TX no separate blocking step, but done with primary incubation in 3% BSA suggestions: use less antibody (secondary, maybe also primary), include separate blocking step, omit pre-extraction step)

Abcam community

Verified customer

Asked on Nov 15 2011

Answer

Thank you for contacting us. I am sorry to hear that the antibody is giving problems. As we discussed yesterday over the phone, the following suggestions might help to get rid of the red blob you are seeing in the nuclei.: 1) use less antibody (for the secondary: e.g. 1/500 or even 1/1000, maybe also for the primary: e.g. 1/1000) 2) include a separate blocking step (e.g. 3-5% BSA or 10% goat serum) 3) omit the pre-extraction step Please let me know if these if these suggestions are of help or not. If the tips do not help, please let me know your PO or order number. If the product was purchased within the last 6 months, we can discuss a free of charge replacement, credit or refund according to our Abpromise guarantee. I wish you good luck with your experiments and look forward to hear back from you.

Abcam Scientific Support

Answered on Nov 15 2011

1-10 of 20 Abreviews or Q&A

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Key features and details

Overview

Product name

Description

Host species

Specificity

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Myeloma

Isotype

Light chain type

Research areas

Associated products

Compatible Secondaries

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (69)

Customer reviews and Q&As

Western blot abreview for Anti-Rad51 antibody [14B4]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad51 antibody [14B4]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Rad51 antibody [14B4]

Post-translational
modifications