Heterogeneous Nuclear Ribonucleoprotein U (Scaffold Attachment Factor A) (HNRNPU) antibody

Details for Product No. ABIN319470
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Antigen
Synonyms AA408410, AI256620, AL024194, AL024437, AW557595, C86794, Hnrpu, SAFA, Sp120, hnRNP U, SN1, HNRPU, SAF-A, U21.1, saf-a
Reactivity
Human, Monkey
(40), (21), (18), (14), (13), (3), (1), (1)
Host
Mouse
(29), (11)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
ELISA, Immunocytochemistry (ICC), Immunofluorescence (IF), Immunoprecipitation (IP), Western Blotting (WB)
(30), (11), (10), (8), (8), (7), (4), (2), (2)
Pubmed 1 reference available
Quantity 50 μL
Shipping to United States (Change)
Availability Will be delivered in 6 to 8 Business Days
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Catalog No. ABIN319470
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Immunogen Recombinant full length protein (human) eluted from an oligo (dt) cellulose column.
Clone 3G6
Isotype IgG1
Specificity Human hnRNP-U. Detects a band of approximately 120 kDa. Cross reactivity with monkey.
Purification Protein A Chromatography.
Alternative Name hnRNP-U / HNRNPU
Background The hnRNPs are RNA binding proteins and they complex with heterogeneous nuclear RNA (hnRNA). These proteins are associated with pre-mRNAs in the nucleus and appear to influence pre-mRNA processing and other aspects of mRNA metabolism and transport. While all of the hnRNPs are present in the nucleus, some seem to shuttle between the nucleus and the cytoplasm. The HNRNP proteins have distinct nucleic acid binding properties. hnRNP U is thought to be involved in the packaging of hnRNA into large ribonucleoprotein complexes. During apoptosis, this protein is cleaved in a caspase- dependent way.
Alternate names: HNRPU, Heterogeneous nuclear ribonucleoprotein U, SAFA, Scaffold attachment factor A, U21.1, p120, pp120
Gene ID 3192
NCBI Accession NP_004492
UniProt Q00839
Application Notes ELISA. Western blot. Immunoprecipitation. Immunofluorescence. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol Immunofluorescence protocol - Formaldehyde fixationCollect cells from T. c. unit and remove media from petri dish using suction. Wash with 1x PBS and remove. Incubate cells in pre-warm (37°C) Para-Formaldehyde for 12 minutes at room temperatureon an orbital shaker. Remove PFA and incubate in 0. 5% Triton X-IOO in 1x PBS for 5 minutes at roomtemperature. Prepare blocking reagent, this is also the antibody diluent. Wash cells 2x with 1x PBS at room temperature, for 4 minutes/wash on an orbital shaker. Block with 1 % NCS and 1x PBS for 30 minutes at room temperature. Prepare primary antibodies (50μl/coverslip) and moist staining chambers. Wash cells 2x with lx PBS at room temperature and air dry briefly. Incubate with primary antibody for 1 hr at room temperature in the dark in stainingchambers. During this time prepare the secondary antibody. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)Incubate with secondary antibody for 1 hour at room temperature in the dark in stainingchambers. Wash cells 5x with 1x PBS. Mount in Dapi. Solutions (prepare fresh the same day of staining): 1x Phosphate buffered saline. Blocking reagent: 1% NCS in 1x PBS (use fresh l0x PBS). Fixation solution: 3. 5% Para formaldehyde. 1. 75g PFA in 20 ml d. H20 plus 5 drops 1M NaOH. Stir on a hot plate at 50-60°C untildissolved. Add 4 drops IN HCI and check pH indicator strip. PH should be 7. 4. Completevolume with d. H20 to 25ml and add 25ml 2xPBS. Check pH before adding to cover slips. Immunofluorescence protocol - Methanol/acetone fixationCollect cells from T. C. unit and remove media from petri dish using suction. Wash with 1x PBS and remove. Fix cells with cold methanol: acetone 1: 1 for 10 minutes on ice. Prepare blocking reagent, this is also the diluent for the antibodies. Remove fixative and wash cells 3x with Ix PBS at RT, for 4 minutes/wash on orbital shaker. Block with 1% NCS and Ix PBS for 30 minutes at RT. Prepare primary antibodies (50μl/coverslip) and moist staining chambers. Wash cells 2x with 1 x PBS at RT and air dry for approximately 7 minutes. Incubate with primary antibody for 1 hr at RT in the dark in staining chambers. During thistime prepare secondary antibody. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)Incubate with secondary antibody for 1 hr at R T in the dark in staining chambers. Wash cells 5x with 1x PBS. Mount in Dapi. Solutions (prepare fresh the same day of staining): 1x Phosphate buffered saline. Blocking reagent: 1% NCS in 1x PBS (use fresh 10x PBS). Fixation solution: methanol: acetone 1: 1 ice cold. Western Blotting ProtocolTransfer gel to PDVF or nitrocellulose membranePlace membrane in plastic tray in blocking buffer for one hour with agitationRinse in wash bufferIncubate in wash buffer plus primary antibody for one hourWash 6 X 5 minutes with wash bufferIncubate in wash buffer plus secondary antibody for one hour
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer PBS with 0.09% sodium azide as preservative.
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 freezing and thawing.
Storage -20 °C
Expiry Date 12 months
Background publications Dreyfuss, Choi, Adam: "Characterization of heterogeneous nuclear RNA-protein complexes in vivo with monoclonal antibodies." in: Molecular and cellular biology, Vol. 4, Issue 6, pp. 1104-14, 1984 (PubMed).

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