RHBDF2 anticorps (N-Term)

N° du produit ABIN657511

Cet anticorps Lapin Polyclonal validé indépendamment détecte spécifiquement RHBDF2 dans WB et IHC (p). Il présente une réactivité avec des échantillons de Humain et Souris.

Aperçu rapide pour RHBDF2 anticorps (N-Term) (ABIN657511)

Antigène: RHBDF2 (Rhomboid 5 Homolog 2 (RHBDF2))

Reactivité: Humain, Souris

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp RHBDF2 est non-conjugé

Application: Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Clone: RB32834

Détail du produit anti-RHBDF2 anticorps

Épitope: AA 80-109, N-Term

Purification: This antibody is purified through a protein A column, followed by peptide affinity purification.

Immunogène: This RHBDF2 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 80-109 amino acids from the N-terminal region of human RHBDF2.

Isotype: IgG

Information d'application

Indications d'application: WB: 1:2000. WB: 1:2000. WB: 1:1000. IHC-P: 1:25. IHC-P: 1:25

Restrictions: For Research Use only

Independent Validation (WB)

Validation #104634 (Western Blotting)

by: Prof. Ludwig

No.: #104634

Date: 19.01.2026

Antigène: RHBDF2

Numéro du lot: SA180724YE

Application validée: Western Blotting

Contrôle positif:

Overexpressed human and murine iRhom2.

Contrôle négative:

HEK293 iRhom1/iRhom2 double knockout.

Conclusion:

The iRhom2 antibody ABIN2846535 works in WB at a 1:1000 dilution to detect overexpressed human iRhom2. Non-specific bands are also detected.

Validation Images

Western blot using ABIN2846535 to reveal iRhom2 (full length 110 kDa, cleaved 55 kDa) in lysates from HEK293 cells (1), HEK293 iRhom1/iRhom2 double knockout (dKO) (2), HEK293 iRhom1/iRhom2 dKO + overexpression of human iRhom2 wt (3), HEK293 iRhom1/iRhom2 dKO + overexpression of murine iRhom2 wt (4)

Protocole

Anticorps primaire: ABIN2846535

Full Protocol:

• Lyse HEK293 cells with lysis buffer (50 mM Tris, 137 mM NaCl, 2 mM EDTA, and 10 mM 1,10-Phenanthroline, adjusted to pH 7.5 and supplemented with 1% Triton X-100 (Sigma-Aldrich) and cOmplete™ protease inhibitor cocktail (Sigma-Aldrich)) for 40 min at 4°C.
• Clear lysates by centrifugation at 20,000 rpm for 20 min at 4°C.
• Denature samples with reducing Lämmli buffer (3% (w/v) SDS, 16% glycerol, 8% 2-mercaptoethanol, 0.01% (w/v) bromophenol blue, and 0.1 M Tris-HCl at pH 6.8) for 20 min at 60°C.
• Separate protein samples on a denaturing 12% SDS-PAGE gel alongside a PageRuler™ Plus Prestained Protein Ladder (Thermo Fisher, 26619).
• Wet the Immobilon®-FL PVDF membrane (0.45 µm pore size) (Millipore, IPFL00010) in methanol for 30 sec.
• Wash membrane once with Milli-Q® water for 1 min.
• Electro-transfer proteins onto Immobilon®-FL PVDF membrane (0.45 µm pore size) (Millipore, IPFL00010) at 100 V for 2 h, cooled on ice.
• Air dry the blot.
• Wet the Immobilon®-FL PVDF membrane again in methanol for 30 sec.
• Wash membrane once with Milli-Q® water for 1 min.
• Block membrane with TBST (TBS + 0.1% Tween-20; TBS = 50 mM Tris, 150 mM NaCl, pH 7.4) containing 5% non-fat dry milk for 20 min at room temperature.
• Wash membrane 3× for 10 min with TBST.
• Incubate membrane with primary rabbit anti-iRhom2 antibody (antibodies-online, ABIN2846535, lot SA180724YE) diluted 1:1,000 in TBST containing 1% BSA overnight at 4°C.
• Wash membrane 3× for 10 min with TBST.
• Incubate membrane with secondary donkey anti-rabbit IgG (H+L), highly cross-adsorbed, Alexa Fluor™ Plus 800 (Thermo Fisher, A32808) diluted 1:100,000 in TBST containing 1% BSA for 1 h at room temperature.
• Wash membrane 1× for 10 min with TBST and 2× for 10 min with TBS.
• Detect stained proteins via fluorescence using the Odyssey 9120 imager system (LI-COR).

Notes:

• Human or murine iRhom2 (lanes 3 and 4) were overexpressed using the pMOWS vector via stable transduction with retroviruses produced by Phoenix Ampho cells, as described in Düsterhöft et al. 2021.
• iRhom1/iRhom2 double-knockout HEK293 cells were generated using the CRISPR-Cas9 system.
• No endogenous human iRhom2 was detected in the lysate of HEK293 cells (lane 1).
• Human iRhom2 was successfully detected in iRhom1/iRhom2 double-knockout HEK293 cells overexpressing human iRhom2 (lane 3).
• Overexpressed murine iRhom2 in iRhom1/iRhom2 double-knockout HEK293 cells was detected at substantially lower levels than overexpressed human iRhom2 (lane 4).
• Non-specific bands were also detected.

Stockage

Format: Liquid

Buffer: Purified polyclonal antibody supplied in PBS with 0.09 % (W/V) 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: 4 °C,-20 °C

Stockage commentaire: RHBDF2 Antibody (N-term) can be refrigerated at 2-8 °C for up to 6 months. For long term storage, place the at -20 °C.

Date de péremption: 6 months

Détails sur RHBDF2

Antigène: RHBDF2 (Rhomboid 5 Homolog 2 (RHBDF2))

Autre désignation: RHBDF2

Sujet: Not expected to have protease activity (By similarity).

Poids moléculaire: 96686

ID gène: 79651

NCBI Accession: NP_001005498, NP_078875

UniProt: Q6PJF5

Pathways: Growth Factor Binding