ARHGEF4 antibody (N-Term)

Catalog No. ABIN2787993

Product Details anti-ARHGEF4 Antibody

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

Binding Specificity: N-Term

Reactivity: Human, Mouse, Rat, Horse, Rabbit, Cow, Dog, Guinea Pig

Host: Rabbit

Clonality: Polyclonal

Conjugate: This ARHGEF4 antibody is un-conjugated

Application: Western Blotting (WB)

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

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

Application Details

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

Comment:

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: 12/23/2016

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.

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.

Full Methods

Primary Antibody: ABIN2787993

Secondary Antibody: 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.

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

Handling

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.

Target Details for ARHGEF4

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

Alternative Name: Arhgef4 (ARHGEF4 Products)

Synonyms: ARHGEF4 antibody, ASEF antibody, ASEF1 antibody, GEF4 antibody, STM6 antibody, 9330140K16Rik antibody, Asef antibody, ENSMUSG00000070955 antibody, Rho guanine nucleotide exchange factor 4 antibody, uncharacterized LOC100470646 antibody, rho guanine nucleotide exchange factor 4 antibody, Rho guanine nucleotide exchange factor (GEF) 4 antibody, ARHGEF4 antibody, arhg4 antibody, LOC100470646 antibody, LOC100544917 antibody, Arhgef4 antibody

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

Pathways: Neurotrophin Signaling Pathway