RHOT1 Antikörper (N-Term)

Produktnummer ABIN405704

Produktdetails anti-RHOT1 Antikörper

Target: RHOT1 (Ras Homolog Gene Family, Member T1 (RHOT1))

Bindungsspezifität: N-Term

Reaktivität: Human, Maus, Ratte, Hund, Rind (Kuh), Meerschweinchen, Pferd, Kaninchen, Zebrafisch (Danio rerio)

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser RHOT1 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB)

Sequenz: EMKPACIKAL TRIFKISDQD NDGTLNDAEL NFFQRICFNT PLAPQALEDV

Homologie: Cow: 100%, Dog: 100%, Guinea Pig: 100%, Horse: 100%, Human: 100%, Mouse: 100%, Rabbit: 100%, Rat: 100%, Zebrafish: 85%

Produktmerkmale: This is a rabbit polyclonal antibody against RHOT1. It was validated on Western Blot using a cell lysate as a positive control.

Aufreinigung: Affinity Purified

Immunogen: The immunogen is a synthetic peptide directed towards the N terminal region of human RHOT1

Anwendungsinformationen

Applikationshinweise: Optimal working dilutions should be determined experimentally by the investigator.

Kommentare:

Antigen size: 659 AA

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #100008 (Western Blotting)

by: Shaw Lab, Department of Biochemistry, University of Utah

No.: #100008

Datum: 12.09.2015

Antigen: RHOT1

Validierte Anwendung: Western Blotting

Positivkontrolle:

Purified recombinant Miro1

Isolated mitochondria from immortalized Miro1 KO MEFs expressing Myc-Miro1

Negativkontrolle:

Purified recombinant Miro2

Isolated mitochondria from immortalized Miro1 WT MEFs

Isolated mitochondria from immortalized Miro1 KO MEFs expressing Myc-Miro2

Bewertung:

Passed. ABIN405704 specifically recognizes Miro1 without cross-reaction with Miro2.

Validierungsbilder

Protokoll

Primärantikörper: ABIN405704

Sekundärantikörper: Goat-anti-rabbit IgG HRP conjugated (Sigma, A0545-1M22)

Full Protocol:

• Transfect immortalized Miro1-/- MEFs with Myc-Miro1-V1 or Myc-Miro2-V1 as described in PMID 25136135.
• Isolate mitochondria from immortalized Miro1+/+, Miro1-/-, Miro1-/- + Myc-Miro1 and Miro1-/- + Myc-Miro2 MEFs.
• Determine protein concentration and snap freeze mitochondria in liquid nitrogen.
• Thaw 30 µg mitochondria per lane on ice.
• Thaw 100 ng recombinant Miro1 and Miro2 per lane on ice.
• Add 2x Laemmli buffer and 2% BME to samples and denature for 5 min at 95 °C.
• Separte 30 µg/lane mitochondria and 100 ng/lane recombinant protein on a 12.5% SDS-PAGE for 3 h at 40mA/180V.
• Semi-dry transfer to PVDF membrane for 2 h at 250 mA/15 V.
• Block the membrane with TBST + 5% non-fat dry milk for 1 h at RT.
• Incubate with primary
• • rabbit anti-RHOT1 antibody (antibodies-online, ABIN405704) diluted 1:1000 in TBST + 3% BSA overnight at 4°C.
  • rabbit anti-Myc1 antibody diluted 1:1000 in TBST + 3% BSA overnight at 4°C.
  • rabbit anti-Cyt c antibody diluted 1:1000 in TBST + 3% BSA overnight at 4°C.
• Wash membrane 3x for 5 min with TBST buffer.
• Incubate with secondar goat anti-rabbit IgG HRP antibody (Sigma, A0545-1M2) diluted 1:2000 in TBST + 5% non-fat dry milk for 1 h at RT.
• Wash membrane 3x for 10 min with TBST buffer.
• Reveal protein bands on a Bio-Rad imaging system using homemade ECL prepared according to PMID 17141265.
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Handhabung

Format: Liquid

Konzentration: Lot specific

Buffer: Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.

Konservierungsmittel: Sodium azide

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

Handhabung: Avoid repeated freeze-thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: For short term use, store at 2-8°C up to 1 week. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.

Details zu RHOT1

Target: RHOT1 (Ras Homolog Gene Family, Member T1 (RHOT1))

Andere Bezeichnung: RHOT1 (RHOT1 Produkte)

Synonyme: rhot1 antikoerper, zgc:55581 antikoerper, zgc:77063 antikoerper, wu:fd14e11 antikoerper, wu:fi14a05 antikoerper, arht2 antikoerper, miro-2 antikoerper, rasl antikoerper, ARHT1 antikoerper, MIRO-1 antikoerper, MIRO1 antikoerper, 2210403N23Rik antikoerper, AA415293 antikoerper, AF244542 antikoerper, AI834919 antikoerper, Arht1 antikoerper, C430039G08Rik antikoerper, Miro1 antikoerper, miro1 antikoerper, si:dkeyp-97g3.6 antikoerper, ras homolog family member T1a antikoerper, ras homolog family member T2 antikoerper, ras homolog family member T1 antikoerper, ras homolog family member T1 L homeolog antikoerper, rhot1a antikoerper, rhot2 antikoerper, RHOT1 antikoerper, rhot1 antikoerper, rhot1.L antikoerper, Rhot1 antikoerper, rhot1b antikoerper

Hintergrund: RHOT1 is mitochondrial GTPase involved in mitochondrial trafficking. It is probably involved in control of anterograde transport of mitochondria and their subcellular distribution.
Alias Symbols: ARHT1, FLJ11040, FLJ12633, MIRO-1, MIRO1
Protein Interaction Partner: PCSK9, ERLIN2, PARK2, ILK, UBC, CFTR, PINK1, UBD, IRAK1, AMFR,
Protein Size: 659

Molekulargewicht: 75 kDa

Gen-ID: 55288

NCBI Accession: NM_001033566, NP_001028738

UniProt: Q8IXI2