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Ordering Information
- : AB3756
- : 100 µg
Anti-Rad51
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M | WB, IC, IH | Rabbit | Affinity Purified | Polyclonal Antibody |
Description:
Anti-Rad51
Trade Name:
Chemicon (Millipore)
Specificity:
Recognizes RAD51. The molecular weight of the protein is 36,963 Daltons.
Immunogen:
Synthetic peptide, amino acids 25-41 of RAD51.
Species Reactivity:
- Human
- Mouse
Application Notes:
Western blot: 1:200-1:1,000 using ECL.
Immunohistochemistry: 1:200-1:1,000.
Immunocytochemistry: 1:200-1:1,000.
Optimal working dilutions must be determined by the end user.
SUGGESTED PROTOCOL FOR IMMUNOHISTOCHEMISTRY
1. Dissect tissues and freeze on dry ice
2. Cut on a cryostat - 10um slices
3. Dry slides at room temperature for 30-90 min
4. Fix with cold Acetone/Methanol (50/50) 2 min - then either process further or store at -20°C. Alternative is fixation with 4% PFA and treatment with a trypsin solution. (0.05% trypsin, incubation 10 minutes at 37°C, followed by 3 washes with PBS)
5. Let air dry
6. PBS 5min
7. Incubate in 50mM ammonium chloride 30 min
8. PBS 5min
9. Blocking serum 30-45min
10. Primary antibody 90min - dilution 1:100 - 1:600
11. Wash 3 times for 5 min
12. Secondary fluorescent conjug. antibody 30min
13. Wash 3 times for 5 min and mount
Immunohistochemistry: 1:200-1:1,000.
Immunocytochemistry: 1:200-1:1,000.
Optimal working dilutions must be determined by the end user.
SUGGESTED PROTOCOL FOR IMMUNOHISTOCHEMISTRY
1. Dissect tissues and freeze on dry ice
2. Cut on a cryostat - 10um slices
3. Dry slides at room temperature for 30-90 min
4. Fix with cold Acetone/Methanol (50/50) 2 min - then either process further or store at -20°C. Alternative is fixation with 4% PFA and treatment with a trypsin solution. (0.05% trypsin, incubation 10 minutes at 37°C, followed by 3 washes with PBS)
5. Let air dry
6. PBS 5min
7. Incubate in 50mM ammonium chloride 30 min
8. PBS 5min
9. Blocking serum 30-45min
10. Primary antibody 90min - dilution 1:100 - 1:600
11. Wash 3 times for 5 min
12. Secondary fluorescent conjug. antibody 30min
13. Wash 3 times for 5 min and mount
Presentation:
Affinity purified immunoglobulin. Precipitated antibody in a solution of 50% saturated ammonium sulfate and PBS containing no preservatives.
Storage Conditions:
Maintain unopened vial at -70C for up to 6 months. Avoid repeated freeze/thaw cycles.
Preparation & Use: To reconstitute the antibody, centrifuge the antibody vial at moderate speed (5,000 rpm) for 5 minutes to pellet the precipitated antibody product. Carefully remove the ammonium sulfate/PBS buffer solution and discard. It is not necessary to remove all of the ammonium sulfate/PBS solution: 10 μL of residual ammonium sulfate solution will not effect the resuspension of the antibody. Do not let the protein pellet dry, as severe loss of antibody reactivity can occur.
Resuspend the antibody pellet in any suitable biological buffer, standard PBS or TBS (pH 7.3-7.5) are typical. Volumes required are not critical but it is suggested that the final antibody concentration be between 0.1 mg/mL and 1.0 mg/mL. For example, to achieve a 1 mg/mL concentration with 50 μg of precipitated antibody, the amount of buffer needed would be 50 μL.
Carefully add the liquid buffer to the pellet. DO NOT VORTEX. Mix by gentle stirring with a wide pipet tip or gentle finger-tapping. Let the precipitated antibody rehydrate for 1 hour at 4-25C° prior to use. Small particles of precipitated antibody that fail to resuspend are normal. Vials are overfilled to compensate for any losses.
The rehydrated antibody solutions can be stored undiluted at 2-8C° for 2 months without any significant loss of activity. Note, the solution is not sterile, thus care should be taken if product is stored at 2-8C°. For storage at -20C°, the addition of an equal volume of glycerol can be used, however, it is recommended that ACS grade or higher glycerol be used, as significant loss of activity can occur if the glycerol used is not of high quality.For freezing, it is recommended that the rehydrated antibody solution be further diluted 1:1 with a 2% BSA (fraction V, highest-grade available) solution made with the rehydration buffer. The resulting 1% BSA/antibody solution can be aliquoted and stored frozen at -70C° for up to 6 months. Avoid repeated freeze/thaw cycles.
Preparation & Use: To reconstitute the antibody, centrifuge the antibody vial at moderate speed (5,000 rpm) for 5 minutes to pellet the precipitated antibody product. Carefully remove the ammonium sulfate/PBS buffer solution and discard. It is not necessary to remove all of the ammonium sulfate/PBS solution: 10 μL of residual ammonium sulfate solution will not effect the resuspension of the antibody. Do not let the protein pellet dry, as severe loss of antibody reactivity can occur.
Resuspend the antibody pellet in any suitable biological buffer, standard PBS or TBS (pH 7.3-7.5) are typical. Volumes required are not critical but it is suggested that the final antibody concentration be between 0.1 mg/mL and 1.0 mg/mL. For example, to achieve a 1 mg/mL concentration with 50 μg of precipitated antibody, the amount of buffer needed would be 50 μL.
Carefully add the liquid buffer to the pellet. DO NOT VORTEX. Mix by gentle stirring with a wide pipet tip or gentle finger-tapping. Let the precipitated antibody rehydrate for 1 hour at 4-25C° prior to use. Small particles of precipitated antibody that fail to resuspend are normal. Vials are overfilled to compensate for any losses.
The rehydrated antibody solutions can be stored undiluted at 2-8C° for 2 months without any significant loss of activity. Note, the solution is not sterile, thus care should be taken if product is stored at 2-8C°. For storage at -20C°, the addition of an equal volume of glycerol can be used, however, it is recommended that ACS grade or higher glycerol be used, as significant loss of activity can occur if the glycerol used is not of high quality.For freezing, it is recommended that the rehydrated antibody solution be further diluted 1:1 with a 2% BSA (fraction V, highest-grade available) solution made with the rehydration buffer. The resulting 1% BSA/antibody solution can be aliquoted and stored frozen at -70C° for up to 6 months. Avoid repeated freeze/thaw cycles.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- RAD51
- RAD51L3
- HsT16930
- BRCC5
- hRAD51
- HRAD51
- RECA
- HsRad51
- RAD51D
- HsRAD51
- RAD51A
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Entrez Gene Summary:
The protein encoded by this gene is a member of the RAD51 protein family. RAD51 family members are highly similar to bacterial RecA and Saccharomyces cerevisiae Rad51, and are known to be involved in the homologous recombination and repair of DNA. This protein can interact with the ssDNA-binding protein RPA and RAD52, and it is thought to play roles in homologous pairing and strand transfer of DNA. This protein is also found to interact with BRCA1 and BRCA2, which may be important for the cellular response to DNA damage. BRCA2 is shown to regulate both the intracellular localization and DNA-binding ability of this protein. Loss of these controls following BRCA2 inactivation may be a key event leading to genomic instability and tumorigenesis. Two alternatively spliced transcript variants of this gene, which encode distinct proteins, have been reported. Transcript variants utilizing alternative polyA signals exist.
Key Applications:
- Western Blotting
- Immunocytochemistry
- Immunohistochemistry
UniProt Summary:
FUNCTION: SwissProt: Q06609 # May participate in a common DNA damage response pathway associated with the activation of homologous recombination and double-strand break repair. Binds to single and double stranded DNA and exhibits DNA-dependent ATPase activity. Underwinds duplex DNA and forms helical nucleoprotein filaments.
SIZE: 339 amino acids; 36966 Da
SUBUNIT: Interacts with BRCA1, BRCA2 and either directly or indirectly with p53. Interacts with XRCC3, RAD54L and RAD54B. Part of a complex with RAD51C and RAD51B. Interacts with RAD51AP1. Interacts with CHEK1/CHK1, and this may require prior phosphorylation of CHEK1.
SUBCELLULAR LOCATION: Nucleus. Note=Colocalizes with RAD51AP1 to multiple nuclear foci upon induction of DNA damage.
TISSUE SPECIFICITY: Highly expressed in testis and thymus, followed by small intestine, placenta, colon, pancreas and ovary. Weakly expressed in breast.
PTM: Phosphorylated. Phosphorylation of Thr-309 by CHEK1/CHK1 may enhance association with chromatin at sites of DNA damage and promote DNA repair by homologous recombination.DISEASE:SwissProt: Q06609 # Defects in RAD51 are associated with breast cancer (BC) [MIM:114480].
SIMILARITY: SwissProt: Q06609 ## Belongs to the recA family. RAD51 subfamily. & Contains 1 HhH domain.
SIZE: 339 amino acids; 36966 Da
SUBUNIT: Interacts with BRCA1, BRCA2 and either directly or indirectly with p53. Interacts with XRCC3, RAD54L and RAD54B. Part of a complex with RAD51C and RAD51B. Interacts with RAD51AP1. Interacts with CHEK1/CHK1, and this may require prior phosphorylation of CHEK1.
SUBCELLULAR LOCATION: Nucleus. Note=Colocalizes with RAD51AP1 to multiple nuclear foci upon induction of DNA damage.
TISSUE SPECIFICITY: Highly expressed in testis and thymus, followed by small intestine, placenta, colon, pancreas and ovary. Weakly expressed in breast.
PTM: Phosphorylated. Phosphorylation of Thr-309 by CHEK1/CHK1 may enhance association with chromatin at sites of DNA damage and promote DNA repair by homologous recombination.DISEASE:SwissProt: Q06609 # Defects in RAD51 are associated with breast cancer (BC) [MIM:114480].
SIMILARITY: SwissProt: Q06609 ## Belongs to the recA family. RAD51 subfamily. & Contains 1 HhH domain.
Qty/Pk:
100 µg
Brand Family:
Chemicon
Format:
Affinity Purified
Host:
Rabbit
Product Name:
Anti-Rad51
Antibody Type:
Polyclonal Antibody