rho Guanine Nucleotide Exchange Factor (GEF) 4 (ARHGEF4) (N-Term) antibody

Details for Product No. ABIN2787993, Supplier: Log in to see
Antigen
  • ARHGEF4
  • ASEF
  • ASEF1
  • GEF4
  • STM6
  • 9330140K16Rik
  • Asef
  • ENSMUSG00000070955
  • Rho guanine nucleotide exchange factor (GEF) 4
  • Rho guanine nucleotide exchange factor 4
  • ARHGEF4
  • arhg4
  • Arhgef4
Alternatives
anti-Human rho Guanine Nucleotide Exchange Factor (GEF) 4 antibody for Western Blotting
Epitope
N-Term
12
9
9
8
5
1
1
1
1
Reactivity
Cow (Bovine), Dog (Canine), Guinea Pig, Horse (Equine), Human, Mouse (Murine), Rabbit, Rat (Rattus)
46
1
Host
Rabbit
20
16
10
Clonality
Polyclonal
Conjugate
Un-conjugated
7
3
3
3
3
3
Application
Western Blotting (WB)
42
29
9
2
1
Options
Supplier
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Available images

'Independent Validation' Badge
Antigen ARHGEF4
Lot Number QC30159-41213
Method validated Western Blotting
Positive Control primary mouse microglia
Notes Passed. ABIN2787993 specifically recognizes Arhgef4 in primary mouse microglia lysates.
Primary Antibody ABIN2787993
Secondary Antibody anti-rabbit HRP-conjugated antibodies (DakoCytomation, P0448, lot 00055814)
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.
Experimental Notes The ARHGEF4 antibody ABIN2787993 reveals multiple bands as is expected for murine Arhgef4.
Validation Images
Western Blotting validation image for anti-rho Guanine Nucleotide Exchange Factor (GEF) 4 (ARHGEF4) (N-Term) antibody (ABIN2787993) Indicated amounts of total protein from primary mouse microglia cell lysate were sepa...
Immunogen The immunogen is a synthetic peptide directed towards the N-terminal region of Mouse Arhgef4
Blocking Peptide Blocking peptide for this product available: ABIN1541270
Sequence ADDYEAPRAG AREADDSGPE AQSCKDQMRT NVINEILSTE RDYIKHLRDI
Predicted Reactivity Cow: 79%, Dog: 93%, Guinea Pig: 92%, Horse: 86%, Human: 100%, Mouse: 86%, Rabbit: 100%, Rat: 100%
Characteristics This is a rabbit polyclonal antibody against Arhgef4. It was validated on Western Blot.
Purification Affinity Purified
Alternative Name Arhgef4 (ARHGEF4 Antibody Abstract)
Background 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
Molecular Weight 57 kDa
Gene ID 226970
NCBI Accession NM_183019, NP_898840
UniProt Q7TNR9
Research Area Signaling, Receptors, Proteolysis / Ubiquitin, Immunology, Cytokines, Growth Factors, Inflammation
Pathways Neurotrophin Signaling Pathway
Application Notes Optimal working dilutions should be determined experimentally by the investigator.
Comment

Antigen size: 484 AA

Restrictions For Research Use only
Format Liquid
Concentration Lot specific
Buffer Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment 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.
Supplier Images
Western Blotting (WB) image for anti-rho Guanine Nucleotide Exchange Factor (GEF) 4 (ARHGEF4) (N-Term) antibody (ABIN2787993) Host: Rabbit
Target Name: Arhgef4
Sample Tissue: Mouse Testis lysates <...