RAB11A, Member RAS Oncogene Family (RAB11A) (C-Term) Antikörper

Details zu Produkt Nr. ABIN2346690, Anbieter: Anmelden zum Anzeigen
Antigen
  • rab11
  • RAB11A
  • wu:fi15h09
  • zgc:103679
  • yl8
  • Rab11a
  • rab11A
  • DDBDRAFT_0190819
  • DDBDRAFT_0191190
  • DDB_0190819
  • DDB_0191190
  • YL8
  • RAB11
  • RAB11A, member RAS oncogene family S homeolog
  • RAB11A, member RAS oncogene family
  • RAB11a, member RAS oncogene family
  • Rab11 GTPase
  • rab11A protein
  • Rab11a, GTPase
  • Rab GTPase
  • rab11A, RAB family GTPase
  • rab11a.S
  • RAB11A
  • rab11a
  • Rab11a
  • rab11A
Epitop
C-Term
20
10
10
9
8
7
5
4
4
2
2
1
1
1
1
Reaktivität
Human
113
29
22
11
9
9
8
7
6
6
6
5
4
4
3
3
3
2
1
1
1
Wirt
Kaninchen
57
44
17
Klonalität
Polyklonal
Konjugat
Unkonjugiert
8
8
6
4
4
4
2
2
2
2
2
2
2
2
2
2
2
Applikation
Immunofluorescence (IF), Immunoprecipitation (IP), Immunohistochemistry (IHC), ELISA, Western Blotting (WB)
109
49
10
4
3
2
1
1
Optionen
Hersteller
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Hersteller Produkt- Nr.
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'Independent Validation' Siegel
Antigen RAB11A
Chargennummer L16102562
Validierte Anwendung Western Blotting
Positivkontrolle

Cell lysates from total human epidermis

Cell lysates from reconstructed human epidermis transfected with control shRNA

Negativkontrolle

Cell lysates from reconstructed human epidermis transfected with RAB11A shRNA

Bewertung Passed. ABIN2346690 specifically recognizes Rab11a in human epidermal samples.
Primärantikörper ABIN2346690
Sekundärantikörper goat anti-rabbit Ig HRP-conjugated antibody (Southern Biotech, 4030-05, lot J1810)
Protokoll
  • Obtain total human epidermis from abdominal dermolipectomy and treat with dispase (Sigma-Aldrich, D4693) to dissociate it from the dermis.
  • For production of reconstructed human epidermis, obtain isolated keratinocytes from epidermal fragments. Grow cells in Epilife (Invitrogen Life Technologies, MEPICF500, lot 1838688) supplemented with penicillin and streptomycin (Sigma-Aldrich, P4333, lot 1868847), HKGS (Human Keratinocyte Growth Supplement, ThermoFisher Scientific, S0015, lot 1848268), 1.5mM calcium (Sigma-Aldrich 25011), KGF (Keratinocyte Growth Factor, Sigma-Aldrich, K1767, lot MKBX7825V) and vitamin C (Sigma-Aldrich, 49752) on polycarbonate culture inserts (Merck Millipore, PIHP01250) at 37°C and 5% CO2, according to Reynier et al. (2016).
  • For silencing with interfering shRNA, transduce keratinocytes with MISSION pLKO.1 puro vector-based lentiviral particles containing a puromycin resistance gene and an shRNA insert under the human U6 promoter (Sigma-Aldrich, TRCN0000303291 for Rab11A and SHC002 for the negative control) following the manufacturer´s instructions, before cell seeding for production of reconstructed human epidermis.
  • Lyse normal or reconstructed epidermis in 300µL/0.6cm2 1x Laemmli buffer (Tris-HCl 63mM pH6.8, SDS 2%, Glycerol 10%, Bromophenol blue 0.0005%, DTT 1mM, all Sigma-Aldrich) and heat at 95°C for 5min.
  • Separate 5µL of total protein extracts in 1x Laemmli buffer on denaturing 10% polyacrylamide SDS gels in a Mini-Protean Tetra Cell electrophoresis cell system (Bio-Rad , 1658004) at 100V for 90min.
  • Transfer proteins onto Hybond ECL membrane (GE Healthcare, RPN3032D) with a Mini TransBlot Western blotting system (Bio-Rad, 1658029) at 100V for 90 min.
  • Block membrane with 1x PBS containing Tween 20 (Sigma-Aldrich) at 0.05% (PBST) and 5% low fat milk at 3% at RT for 1h.
  • Incubate the membrane with primary rabbit anti-human RAB11A, Member RAS Oncogene Family (RAB11A) (C-Term) antibody (antibodies-online, ABIN2346690, lot L16102562) diluted 1:250 in blocking buffer ON at 4°C.
  • Wash the membrane 3x for 15min with 1x PBST.
  • Incubate the membrane with secondary goat anti-rabbit Ig HRP-conjugated antibody (Southern Biotech, 4030-05, lot J1810) diluted 1:10000 in blocking buffer for 90min at RT.
  • Wash the membrane 3x for 15min with 1x PBST.
  • Reveal protein bands using ECL prime Detection Reagent (GE Healthcare, RPN2238, lot 9838391) on a GBox apparatus (Syngene).
Validierungsbilder
Western Blotting validation image for anti-RAB11A, Member RAS Oncogene Family (RAB11A) (C-Term) antibody (ABIN2346690)

Western blot analysis of reconstructed human epidermis cell lysates with ABIN23466...

Immunogen Synthetic peptide corresponding to the C-terminal hypervariable region of the human Rab11a sequence. Sequence Homology: Rab11b protein-10/17.
Isotyp IgG
Kreuzreaktivität Hund, Human, Kaninchen, Ratte (Rattus)
Kreuzreaktivität (Details) Calculated cross reactivity: Ca Hu Rb Rt
Produktmerkmale Rab11A (RAB11A Member RAS Oncogene Family, Rab-11A, MGC1490, RAB11, Rab-11, Ras-related Protein Rab-11A, Ras-related protein Rab11A, YL8)
Reinigung Purified by immunoaffinity chromatography.
Andere Bezeichnung Rab11A (RAB11A Antibody Abstract)
Pathways Regulation of Cell Size, Thromboxane A2 Receptor Signaling, Regulation of long-term Neuronal Synaptic Plasticity
Applikationshinweise Optimal working conditions should be determined by the investigator.
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Buffer Supplied as a liquid in PBS, pH 7.4, 0.09 % 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 -20 °C
Informationen zur Lagerung -20°C
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