PALB2 antibody (AA 1101-1186)

Catalog No. ABIN670536

Product Details anti-PALB2 Antibody

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

Binding Specificity: AA 1101-1186

Reactivity: Human, Pig

Host: Rabbit

Clonality: Polyclonal

Conjugate: This PALB2 antibody is un-conjugated

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))

Cross-Reactivity: Human, Pig

Predicted Reactivity: Mouse,Rat,Cow

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human PALB2

Isotype: IgG

Application Details

Application Notes: 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: 07/01/2014

Lot Number: 120510

Method validated: Western Blotting

Positive Control: A549 cell extract

Negative Control: c6/36 mosquito cell extract (non-reactive species)

Notes: 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).

Full Methods

Primary Antibody:

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

Secondary Antibody:

• 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).

Experimental Notes: - No challenges noted.

Handling

Format: Liquid

Concentration: 1 μg/μL

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

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

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

Expiry Date: 12 months

Target Details for PALB2

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

Alternative Name: PALB2 (PALB2 Products)

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

Background:

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.

Gene ID: 79728

UniProt: Q86YC2

Pathways: DNA Damage Repair