PALB2 anticorps (AA 1101-1186)

N° du produit ABIN670536

Détail du produit anti-PALB2 anticorps

Antigène: PALB2 (Partner and Localizer of BRCA2 (PALB2))

Épitope: AA 1101-1186

Reactivité: Humain, Porc

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp PALB2 est non-conjugé

Application: Western Blotting (WB), ELISA, Flow Cytometry (FACS), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Réactivité croisée: Humain, Porc

Homologie: Mouse,Rat,Cow

Purification: Purified by Protein A.

Immunogène: KLH conjugated synthetic peptide derived from human PALB2

Isotype: IgG

Information d'application

Indications d'application: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (WB)

Validation #029749 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029749

Date: 01.07.2014

Numéro du lot: 120510

Application validée: Western Blotting

Contrôle positif: A549 cell extract

Contrôle négative: c6/36 mosquito cell extract (non-reactive species)

Conclusion: A single strong band was observed in the positive control at the correct molecular weight. Several faint bands appeared in the negative control, but they were much less intense than the positive control band.

Validation Images

Figure 1. Western blot of lysates from A549 cells (Lane 1) and c6/36 cells (Lane 2) probed with anti-PALB2 (upper panel) or with anti-Actin for loading control (lower panel).

Protocole

Anticorps primaire:

• Antigen: Partner and Localizer of BRCA2 (PALB2) (1:500 dilution)
• Catalog number: ABIN670536
• Lot number: 120510

Anticorps secondaire:

• Antibody: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate (1:20,000 dilution)
• Lot number: L170-6515

Full Protocol:

• 1. Total protein extracts were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5 min prior to loading.
• 2. 35 μg of boiled extracts were loaded and resolved on 8-16% SDS-polyacrylamide gel.
• 3. The Thermo Scientific - Spectra Multicolor Broad Range (Cat # 26634) were used as molecular mass markers.
• 4. Proteins were then transferred onto PVDF membrane by wet transfer and protein transfer was confirmed with Ponceau-S staining.
• 5. The PVDF membrane was incubated with 25 mL of blocking buffer [Tris Buffered Saline, pH 7.4 plus 0.1% TW20 (TBST)] containing 5% (W/V) BSA at room temperature for 1 h.
• 6. The membrane was rinsed with TBST once.
• 7. The membrane was immersed with the protein side up in the primary antibody solution (anti-PALB2; 1:500) in TBST containing 5% (W/V) BSA and incubated for 16 h at 4°C.
• 8. The membrane was rinsed in TBST thrice for 5 min each.
• 9. The membrane was incubated in the HRP-conjugated secondary antibody solution (Goat anti-rabbit IgG-HRP; 1:20,000) in TBST containing 5% (W/V) BSA and incubated for 1 h at room temperature (~26°C) with gentle agitation.
• 10. The membrane was rinsed thrice TBST thrice for 5 min each.
• 11. The membrane was rinsed in TBS twice for 30 s each.
• 12. Signals were detected with ECL-2 Substrate. The blot was scanned for 300 s.
• 13. The membrane was rinsed three times TBST.
• 14. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 min each.
• 15. The membrane was washed in TBST 2 times for 10 min each.
• 16. Repeated Steps 5-12 with the loading control antibody (anti-Actin; 1:6,000) and its matching secondary antibody (Goat anti-rabbit IgG-HRP; 1:20,000).

Notes: - No challenges noted.

Stockage

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Agent conservateur: ProClin

Précaution d'utilisation: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Stock: 4 °C,-20 °C

Stockage commentaire: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Date de péremption: 12 months

Détails sur PALB2

Antigène: PALB2 (Partner and Localizer of BRCA2 (PALB2))

Autre désignation: PALB2 (PALB2 Produits)

Synonymes: anticorps PALB2, anticorps Fancn, anticorps si:ch211-14k19.9, anticorps FANCN, anticorps PNCA3, anticorps 4732427B05, anticorps BC066140, anticorps RGD1304759, anticorps partner and localizer of BRCA2, anticorps partner and localizer of BRCA2 L homeolog, anticorps PALB2, anticorps palb2, anticorps palb2.L, anticorps Palb2

Sujet:

Synonyms: FANCN, PNCA3, Partner and localizer of BRCA2, PALB2

Background: Plays a critical role in homologous recombination repair (HRR) through its ability to recruit BRCA2 and RAD51 to DNA breaks. Strongly stimulates the DNA strand-invasion activity of RAD51, stabilizes the nucleoprotein filament against a disruptive BRC3-BRC4 polypeptide and helps RAD51 to overcome the suppressive effect of replication protein A (RPA). Functionally cooperates with RAD51AP1 in promoting of D-loop formation by RAD51. Serves as the molecular scaffold in the formation of the BRCA1-PALB2-BRCA2 complex which is essential for homologous recombination. Via its WD repeats is proposed to scaffold a HR complex containing RAD51C and BRCA2 which is thought to play a role in HR-mediated DNA repair. Essential partner of BRCA2 that promotes the localization and stability of BRCA2. Also enables its recombinational repair and checkpoint functions of BRCA2. May act by promoting stable association of BRCA2 with nuclear structures, allowing BRCA2 to escape the effects of proteasome-mediated degradation. Binds DNA with high affinity for D loop, which comprises single-stranded, double-stranded and branched DNA structures. May play a role in the extension step after strand invasion at replication-dependent DNA double-strand breaks, together with BRCA2 is involved in both POLH localization at collapsed replication forks and DNA polymerization activity.

ID gène: 79728

UniProt: Q86YC2

Pathways: Réparation de l'ADN