RHBDF2 Antikörper (N-Term)

Produktnummer ABIN657511

Dieser unabhängig validierte Kaninchen Polyklonal Antikörper detektiert spezifisch RHBDF2 in WB und IHC (p). Es zeigt Reaktivität gegenüber Proben von Human und Maus.

Kurzübersicht für RHBDF2 Antikörper (N-Term) (ABIN657511)

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

Reaktivität: Human, Maus

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser RHBDF2 Antikörper ist unkonjugiert

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

Klon: RB32834

Produktdetails anti-RHBDF2 Antikörper

Bindungsspezifität: AA 80-109, N-Term

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

Immunogen: 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.

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB: 1:2000. WB: 1:2000. WB: 1:1000. IHC-P: 1:25. IHC-P: 1:25

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #104634 (Western Blotting)

by: Prof. Ludwig

No.: #104634

Datum: 19.01.2026

Antigen: RHBDF2

Chargennummer: SA180724YE

Validierte Anwendung: Western Blotting

Positivkontrolle:

Overexpressed human and murine iRhom2.

Negativkontrolle:

HEK293 iRhom1/iRhom2 double knockout.

Bewertung:

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

Validierungsbilder

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)

Protokoll

Primärantikörper: 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).

Anmerkungen:

• 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.

Handhabung

Format: Liquid

Buffer: Purified polyclonal antibody supplied in PBS with 0.09 % (W/V) sodium azide.

Konservierungsmittel: Sodium azide

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

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: 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.

Haltbarkeit: 6 months

Details zu RHBDF2

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

Andere Bezeichnung: RHBDF2

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

Molekulargewicht: 96686

Gen-ID: 79651

NCBI Accession: NP_001005498, NP_078875

UniProt: Q6PJF5

Pathways: Growth Factor Binding