Retinoblastoma 1 anticorps (pSer780)

N° du produit ABIN675294

Détail du produit anti-Retinoblastoma 1 anticorps

Antigène: Retinoblastoma 1 (RB1)

Épitope: pSer780

Reactivité: Humain, Rat

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp Retinoblastoma 1 est non-conjugé

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

Réactivité croisée: Humain, Rat

Homologie: Mouse,Dog,Cow,Chicken

Purification: Purified by Protein A.

Immunogène: KLH conjugated synthetic phosphopeptide derived from human Rb around the phosphorylation site of (Ser780)

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 #029795 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029795

Date: 21.08.2014

Antigène: Retinoblastoma 1 (RB1) (pSer780)

Numéro du lot: 120508

Application validée: Western Blotting

Contrôle positif: NIH/3T3 cells irradiated with 25 Joules/m2 UV-C

Contrôle négative: Non-irradiated NIH/3T3 cells

Conclusion: A strong band was observed in the positive control sample at the correct molecular weight. An additional band was also observed in the positive sample at a lower molecular weight which may represent non-specific binding. No bands were observed in the negative control sample.

Validation Images

Figure 1: Western Blot for RB1 pSer780. Grey arrowhead indicates the expected molecular weight of ~105 kDa.

Protocole

Anticorps primaire:

• Antigen: Retinoblastoma 1 (RB1) (pSer780)
• Catalog number: ABIN675294
• Lot number: 120508
• Antibody Dilution: 1:100

Anticorps secondaire:

• Antigen: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate
• Lot number: L170-6515
• Antibody Dilution: 1:10,000

Full Protocol:

• 1. The cell extracts were heated at 95°C for 5 minutes in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol.
• 2. 20 μL of heated culture-media 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 hour.
• 6. The membrane was rinsed with TBST once.
• 7. The membrane was immersed with the protein side up in the primary antibody solution in TBST containing 5% (W/V) BSA and incubated for 20 hours at 4°C.
• 8. The membrane was rinsed in TBST thrice for 5 minutes each.
• 9. The membrane was incubated in the HRP-conjugated secondary antibody solution in TBST containing 5% (W/V) BSA and incubated for 1 hour at room temperature (~26°C) with gentle agitation.
• 10. The membrane was rinsed thrice TBST thrice for 5 minutes each.
• 11. The membrane was rinsed in TBS twice for 30 seconds each.
• 12. Signals were detected with ECL-2 Substrate. The blot was scanned for 50 minutes.
• 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 minutes each.
• 15. The membrane was washed in TBST 2 times for 10 minutes each.
• 16. Repeated Steps 5-12 with the loading control antibody (for Anti-actin) and its matching secondary antibody.

Notes: - No experimental 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

Références pour anticorps anti-Retinoblastoma 1 (ABIN675294)

Béguelin, Rivas, Calvo Fernández, Teater, Purwada, Redmond, Shen, Challman, Elemento, Singh, Melnick: "EZH2 enables germinal centre formation through epigenetic silencing of CDKN1A and an Rb-E2F1 feedback loop." dans: Nature communications, Vol. 8, Issue 1, pp. 877, (2018) (PubMed).

Détails sur Retinoblastoma 1

Antigène: Retinoblastoma 1 (RB1)

Autre désignation: Rb/p105-Rb (RB1 Produits)

Synonymes: anticorps OSRC, anticorps RB, anticorps p105-Rb, anticorps pRb, anticorps pp110, anticorps Rb, anticorps Rb-1, anticorps P105-RB, anticorps PP110, anticorps RB11, anticorps zgc:154147, anticorps RB transcriptional corepressor 1, anticorps RB transcriptional corepressor like 1, anticorps retinoblastoma 1, anticorps RB1, anticorps RBL1, anticorps Rb1, anticorps rb1

Sujet:

Synonyms: RbSer780, OSRC, P105 RB, P105RB, PP105, PP110, pRb, RB 1, RB1, RB1 protein, Retinoblastoma 1 including osteosarcoma, Retinoblastoma 1, Retinoblastoma associated protein, Including osteosarcoma, Osteosarcoma, p105-Rb, Rb, RB_HUMAN, Retinoblastoma suspectibility protein, Retinoblastoma-associated protein, Retinoblastoma related osteosarcoma, Retinoblastoma susceptibility gene.

Background: Nuclear Marker.The protein encoded by this gene is a negative regulator of the cell cycle and was the first tumor suppressor gene found. The encoded protein also stabilizes constitutive heterochromatin to maintain the overall chromatin structure. The active, hypophosphorylated form of the protein binds transcription factor E2F1. Defects in this gene are a cause of childhood cancer retinoblastoma (RB), bladder cancer, and osteogenic sarcoma.

ID gène: 5925

Pathways: Cycle Cellulaire, Intracellular Steroid Hormone Receptor Signaling Pathway, Mitotic G1-G1/S Phases, DNA Replication, Maintenance of Protein Location, Synthesis of DNA, Autophagy