CNST anticorps

N° du produit ABIN5575708

Détail du produit anti-CNST anticorps

Antigène: CNST (Consortin, Connexin Sorting Protein (CNST))

Reactivité: Souris

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp CNST est non-conjugé

Application: Western Blotting (WB)

Fonction: Rabbit polyclonal antibody raised against partial recombinant Cnst.

Séquence: MAPEERRDSE DRVSKETEDY LHSLLERCLK DAEDSLSYED IQDDDSDLLQ DLSPEEASYS LQEDLPPDES TLSLDDLAKK IEIAEAIPAE GLVSILKKRN DTVGSHPAQM QQKPAKRRVR FQEID

Specificité: Specific to recombinant protein GX2268. This antibody detects mC1orf71 protein.

Réactivité croisée: Souris

Immunogène: Recombinant GST fusion protein corresponding to 125 amino acids of mouse Cnst.

Information d'application

Indications d'application: Western Blot (1:1000)
The optimal working dilution should be determined by the end user.

Restrictions: For Research Use only

Independent Validation (WB)

Validation #102674 (Western Blotting)

by: Christoffels Lab, Heart Failure Research Center, Academisch Medisch Centrum Amsterdam

No.: #102674

Date: 14.05.2018

Antigène: CNST

Numéro du lot: BIAD001

Application validée: Western Blotting

Contrôle positif: Whole E13.5 wildtype mouse embryo protein lysate

Contrôle négative: Whole E13.5 mouse embryo protein lysates homozygous for a null Cnst allele. This mouse line was generated using CRISPR/Cas9.

Conclusion:

Passed. ABIN5575708 specifically recognizes Cnst in whole embryo lysates.

Validation Images

Western blot detection of consortin whole E13.5 mouse embryo lysates using ABIN5575708. Each lane represents one whole E13.5 embryo lysate of each specified genotype, with CNST at about 75kDa and control GAPDH at about 35kDa.

Protocole

Anticorps primaire: ABIN5575708

Anticorps secondaire: anti-rabbit HRP-conjugated antibody (GE Healthcare, NA934V, lot 9804671)

Full Protocol:

• Isolate E13.5 mouse embryos in cold PBS and snap freeze with liquid nitrogen.
• Lyse each individual embryo in 300µl of cold RIPA buffer.
• Determine total protein content of the lysates using Pierce™ BCA Protein Assay Kit (Thermo Scientific, 23227, lot NA168207).
• Denature 5µg of total protein for 5min at 95°C in 20µl 2x Laemmli SDS buffer and subsequently separate them on a 10% denaturing acrylamide gel in a BIO-RADMini-PROTEAN 3 chamber for 1h at 180V.
• Transfer proteins onto PVDF membrane (Immobilon-P, Millipore) with a Western blotting system for 45min at 25V.
• Block the membrane with 5% Milk in TBST as blocking buffer for 1-2h at RT.
• Incubation with primary
• • rabbit anti-Consortin antibody (antibodies-online, ABIN5575708, lot BIAD001) diluted 1:2000 in blocking buffer 5% milk ON at 4°C.
  • mouse anti-GAPDH antibody (Santa Cruz, sc-365062, lot I0916) diluted 1:5000 in blocking buffer ON at 4°C.
• Wash membrane 5x for 15min with TBST.
• Incubation with secondary
• • anti-rabbit HRP-conjugated antibody (GE Healthcare, NA934V, lot 9804671) diluted 1:2000 blocking buffer for 1-2h at RT.
  • anti-mouse HRP-conjugated antibody (GE Healthcare, NA931V, lot 9800151) diluted 1:2000 in blocking buffer for 1-2h at RT.
• Wash membrane 5x for 15min with TBST.
• Reveal protein bands using ECL detection kit (GE Healthcare) on an ImageQuant LAS 4000 phosphoimaging unit (GE Healthcare).

Notes:

The Consortin antibody ABIN5575708 reveals a protein of the expected molecular weight of antigen in lysates of whole E13.5 embryos. The protein bands is only visible in the positive but not the negative controls.

Stockage

Format: Liquid

Buffer: In PBS (50 % glycerol, 0.02 % sodium azide)

Agent conservateur: Sodium azide

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

Stock: -20 °C

Stockage commentaire: Store at -20°C.
Aliquot to avoid repeated freezing and thawing.

Détails sur CNST

Antigène: CNST (Consortin, Connexin Sorting Protein (CNST))

Autre désignation: 9630058J23Rik (CNST Produits)

Synonymes: anticorps C1orf71, anticorps PPP1R64, anticorps 9630058J23Rik, anticorps consortin, connexin sorting protein, anticorps CNST, anticorps Cnst

Sujet: Full Gene Name: RIKEN cDNA 9630058J23 gene
Synonyms: MGC31547

ID gène: 226744