ARHGEF4 anticorps (N-Term)

N° du produit ABIN2787993

Cet anticorps anti-ARHGEF4 est un anticorps Lapin Polyclonal détectant ARHGEF4 dans WB. Adapté pour Humain, Souris, Rat, Boeuf (Vache), Chien, Cobaye, Cheval et Lapin. Validé indépendamment pour une utilisation dans Western Blotting.

Aperçu rapide pour ARHGEF4 anticorps (N-Term) (ABIN2787993)

Antigène: ARHGEF4 (rho Guanine Nucleotide Exchange Factor (GEF) 4 (ARHGEF4))

Reactivité: Humain, Souris, Rat, Boeuf (Vache), Chien, Cobaye, Cheval, Lapin

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp ARHGEF4 est non-conjugé

Application: Western Blotting (WB)

Détail du produit anti-ARHGEF4 anticorps

Épitope: N-Term

Séquence: ADDYEAPRAG AREADDSGPE AQSCKDQMRT NVINEILSTE RDYIKHLRDI

Homologie: Cow: 79%, Dog: 93%, Guinea Pig: 92%, Horse: 86%, Human: 100%, Mouse: 86%, Rabbit: 100%, Rat: 100%

Attributs du produit: This is a rabbit polyclonal antibody against Arhgef4. It was validated on Western Blot.

Purification: Affinity Purified

Immunogène: The immunogen is a synthetic peptide directed towards the N-terminal region of Mouse Arhgef4

Information d'application

Indications d'application: Optimal working dilutions should be determined experimentally by the investigator.

Commentaires:

Antigen size: 484 AA

Restrictions: For Research Use only

Independent Validation (WB)

Validation #100106 (Western Blotting)

by: Instituto de Parasitología y Biomedicina López-Neyra

No.: #100106

Date: 23.12.2016

Antigène: ARHGEF4

Numéro du lot: QC30159-41213

Application validée: Western Blotting

Contrôle positif: primary mouse microglia

Conclusion: Passed. ABIN2787993 specifically recognizes Arhgef4 in primary mouse microglia lysates.

Validation Images

Indicated amounts of total protein from primary mouse microglia cell lysate were separated on a denaturing SDS-PAGE gel and incubated with ABIN2787993 as described in the protocol.

Protocole

Anticorps primaire: ABIN2787993

Anticorps secondaire: anti-rabbit HRP-conjugated antibodies (DakoCytomation, P0448, lot 00055814)

Full Protocol:

• Primary mouse microglia are grown in DMEM (Invitrogen), supplemented with 10% foetal bovine serum (Gibco), 10% horse serum and penicillin and streptomycin (Gibco), at 37°C and 5% CO2 in a 12-well dish.
• Lyse cells in 25µl per well cold lysis buffer (10mM Tris-HCl pH 8.0, 150mM NaCl, 1% Nonidet-P40, 1mM EDTA, 10mM NaF, 1mM Na₃VO₄, protease inhibitors (Sigma)).
• Determine total protein content of the lysates using Bradford assay (Bio-Rad, 500-0006, lot 111832).
• Denature 5µg, 10µg and 30µg total protein for 5min at 95°C in 5µl, 10µl and 20µl Laemmli SDS sample buffer and subsequently separate them on a denaturing self-made 10% SDS-PAGE for 1h at 140V.
• Transfer proteins onto PVDF membrane (Pall Life Sciences, 75696G, lot TO3225) with a semi-dry Western blotting system for 50min at 60mA.
• Block the membrane with TBST (TBS, 0.1% Tween) containing 5% milk for 1h at RT.
• Wash membrane 3x 5min with TBST.
• Incubation with primary rabbit anti-ARHGEF4 antibody (antibodies-online, ABIN2787993, lot QC30159-41213) diluted 1:500 in TBST containing 2% BSA ON at 4°C.
• Wash membrane 5x 5min with TBST.
• Incubation with anti-rabbit horseradish peroxidase-conjugated secondary antibodies (DakoCytomation, P0448, lot 00055814) diluted 1:2000 in TBST containing 5% milk for 1h at RT.
• Wash membrane 5x 5min with TBST and 3x 5min with TBS.
• Reveal protein bands using ECL Plus Western Blotting Detection Reagent (GE Healthcare, RPN2132, lot 4645641) with Curix RP2 Plus medical X-ray films (AGFA Health Care, ENKMV, lot 79040074) on an AGFA Health Care Curix60-developer.

Notes: The ARHGEF4 antibody ABIN2787993 reveals multiple bands as is expected for murine Arhgef4.

Stockage

Format: Liquid

Concentration: Lot specific

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

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.

Conseil sur la manipulation: Avoid repeated freeze-thaw cycles.

Stock: -20 °C

Stockage commentaire: 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.

Détails sur ARHGEF4

Antigène: ARHGEF4 (rho Guanine Nucleotide Exchange Factor (GEF) 4 (ARHGEF4))

Autre désignation: Arhgef4

Sujet: Arhgef4 acts as guanine nucleotide exchange factor (GEF) for RHOA, RAC1 and CDC42 GTPases. Binding of APC may activate RAC1 GEF activity. The APC-ARHGEF4 complex seems to be involved in cell migration as well as in E-cadherin-mediated cell-cell adhesion By similarity. It is required for MMP9 up-regulation via the JNK signaling pathway in colorectal tumor cells. Arhgef4 is involved in tumor angiogenesis and may play a role in intestinal adenoma formation and tumor progression.
Alias Symbols: 9330140K16Rik, Asef, ENSMUSG00000070955
Protein Size: 484

Poids moléculaire: 57 kDa

ID gène: 226970

NCBI Accession: NM_183019, NP_898840

UniProt: Q7TNR9

Pathways: Neurotrophin Signaling Pathway