anti-Human NCOA4 antibody for Western Blotting

Recommended NCOA4 Antibody (supplied by: Log in to see )

Antigen
Nuclear Receptor Coactivator 4 (NCOA4) Antibodies
  • zgc:55307
  • zgc:77270
  • NCOA4
  • MGC138109
  • rfg
  • ele1
  • ptc3
  • ara70
  • MGC84996
  • DKFZp469A035
  • DKFZp459B2429
  • DKFZp469A1925
  • ncoa4
  • ARA70
  • ELE1
  • PTC3
  • RFG
  • 1110034E15Rik
  • 4432406M01Rik
  • AI227008
  • Rfg
  • nuclear receptor coactivator 4
  • nuclear receptor coactivator 4 S homeolog
  • Ncoa4
  • ncoa4
  • NCOA4
  • ncoa4.S
Epitope
N-Term
12
11
10
8
8
6
4
4
3
3
2
2
1
1
1
1
1
1
Reactivity
Cow (Bovine), Dog (Canine), Guinea Pig, Horse (Equine), Human, Mouse (Murine), Rabbit, Rat (Rattus)
83
15
12
3
3
3
2
1
1
1
Host
Rabbit
44
39
Clonality
Polyclonal
Conjugate
This NCOA4 antibody is un-conjugated
2
2
2
2
2
2
Application
Western Blotting (WB)
76
39
14
14
10
4
2
1
1
1
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Available images

Catalog No. ABIN2780237
$ 289.00
Plus shipping costs $45.00
Relevance Score ABIN Application Conjugate Host Isotype Epitope Supplier Clonality References Details
2.8724732 ABIN2776441 WB Rabbit C-Term Log in to see Polyclonal 0
2.8724732 ABIN2706652 WB Rabbit Center Log in to see Polyclonal 0
2.8724732 ABIN953616 EIA WB Rabbit Ig Fraction C-Term, AA 495-524 Log in to see Polyclonal 2
2.8724732 ABIN3183343 ELISA IHC WB Rabbit IgG Internal Region Log in to see Polyclonal 0
2.8724732 ABIN521736 IF ELISA WB Mouse IgG1 kappa AA 505-614, partial Log in to see 1B7 1
2.8724732 ABIN521735 IHC (p) ELISA WB Mouse IgG1 kappa AA 505-614, partial Log in to see 1E5 0
2.8724732 ABIN521737 IF IHC (p) ELISA WB Mouse IgG Mix lambda AA 505-614, partial Log in to see 1F11 0
2.8724732 ABIN440536 IP WB Rabbit AA 564-614 Log in to see Polyclonal 0
2.8724732 ABIN1537407 WB Rabbit Ig Fraction AA 496-524, C-Term Log in to see Polyclonal 0
2.8724732 ABIN1539864 WB Mouse IgM Log in to see 439CT10-4-4 0
2.8724732 ABIN394861 ELISA IF WB Mouse IgG1 kappa AA 505-615 Log in to see 1B7 0
2.8724732 ABIN395073 ELISA IF IHC WB Mouse IgG Mix lambda AA 505-615 Log in to see 1F11 0
2.8724732 ABIN1539863 WB Mouse IgM Log in to see 439CT10-4-4 0
2.8724732 ABIN395747 IF WB Mouse IgG2b kappa AA 1-576 Log in to see 2H8 0
2.8724732 ABIN948657 ELISA WB Mouse IgG1 kappa AA 505-614, partial Log in to see 1A2 0
2.8724732 ABIN563529 ELISA WB Mouse IgG1 kappa AA 505-614, partial Log in to see 1A2 0
2.8724732 ABIN4338296 WB Mouse IgG1 Log in to see 2G4 0
2.8724732 ABIN5692970 ELISA WB Rabbit AA 22-170 Log in to see Polyclonal 0
2.8724732 ABIN2995417 WB Mouse IgM Log in to see 0
2.8724732 ABIN2916539 WB Mouse IgM Log in to see 0

General

Antigen Nuclear Receptor Coactivator 4 (NCOA4) Antibodies
Epitope N-Term
(12), (11), (10), (8), (8), (6), (4), (4), (3), (3), (2), (2), (1), (1), (1), (1), (1), (1)
Reactivity Cow (Bovine), Dog (Canine), Guinea Pig, Horse (Equine), Human, Mouse (Murine), Rabbit, Rat (Rattus)
(83), (15), (12), (3), (3), (3), (2), (1), (1), (1)
Host Rabbit
(44), (39)
Clonality
Polyclonal
Conjugate This NCOA4 antibody is un-conjugated
(2), (2), (2), (2), (2), (2)
Application Western Blotting (WB)
(76), (39), (14), (14), (10), (4), (2), (1), (1), (1)
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Product Details anti-NCOA4 Antibody

Target Details NCOA4 Application Details Handling Images
Blocking Peptide Blocking peptide for this product available: ABIN980536
Sequence CLIHQLEYTQ NKDLANQVSV CLERLGSLAL KPEDSTVLLF EADTSALRQT
Predicted Reactivity Cow: 92%, Dog: 93%, Guinea Pig: 100%, Horse: 93%, Human: 100%, Mouse: 100%, Rabbit: 100%, Rat: 100%
Characteristics This is a rabbit polyclonal antibody against Ncoa4. It was validated on Western Blot.
Purification Affinity Purified
Immunogen The immunogen is a synthetic peptide corresponding to a region of Mouse
Plasmids, Primers & others

Target Details NCOA4

Product Details anti-NCOA4 Antibody Application Details Handling Images back to top
Antigen
Alternative Name Ncoa4 (NCOA4 Antibody Abstract)
Background The function of Ncoa4 remains unknown.
Alias Symbols: 1110034E15Rik, 4432406M01Rik, AI227008, ARA70, MGC103382, Rfg
Protein Interaction Partner: Ndn,
Protein Size: 625
Molecular Weight 70 kDa
Gene ID 27057
NCBI Accession NM_001033988, NP_001029160
Pathways Intracellular Steroid Hormone Receptor Signaling Pathway

Application Details

Product Details anti-NCOA4 Antibody Target Details NCOA4 Handling Images back to top
Application Notes Optimal working dilutions should be determined experimentally by the investigator.
Comment

Antigen size: 625 AA

Restrictions For Research Use only

Handling

Product Details anti-NCOA4 Antibody Target Details NCOA4 Application Details Images back to top
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.

Images

Product Details anti-NCOA4 Antibody Target Details NCOA4 Application Details Handling back to top
Supplier Images
Western Blotting (WB) image for anti-Nuclear Receptor Coactivator 4 (NCOA4) (N-Term) antibody (ABIN2780237) WB Suggested Anti-Ncoa4 Antibody Titration: 0.2-1 ug/ml ELISA Titer: 1:312500 Pos...
'Independent Validation' Badge
Antigen Ncoa4
Lot Number QC11327
Method validated Western Blotting
Positive Control HIF overexpression mice, WT mice
Negative Control NCOA4-/- mice
Notes Passed, based on the fact that the antibody picked up the positive control (unpublished data suggests that NCOA4 is highly expressed in an intestinal HIF overexpression model).
Primary Antibody ABIN2780237
Secondary Antibody Anti-rabbit HRP-linked secondary antibody (Cell Signaling, 7074S, lot 26)
Protocol
  • Wild type (WT) or NCOA4 -/- mice were fed normal or iron-deficient diet for 7 days, followed by euthanization and scraping of duodenal epithelium (DE).
  • DE cells were lysed in RIPA buffer and whole cell lysate (WCL) prepared.
  • 50µg total protein from each sample were used for Western blot analysis.
  • Samples were denatured in 5x gel-loading buffer containing beta-marcaptoethanol and separated on a 10% SDS-PAGE gel (Laemmli 1970).
  • Transfer onto Nitrocellulose membrane (Biorad, 1620212, lot 10484060) was done on Trans-Blot SD apparatus (Bio-Rad) following the manufacturer’s protocol.
  • Membranes were blocked with TBS-tween (TBST) solution containing 5% milk for 1h.
  • Incubation with primary antibody overnight at 4°C:
    • NCOA4 antibody ABIN2780237 (lot QC11327) diluted 1:1000 in TBST containing 0.1% BSA.
    • FTH1 antibody (Cell signaling, 3998S; lot 1) diluted in TBST containing 0.1% BSA.
    • GAPDH antibody (Santa Cruz, SC-25778, lot D30115) diluted in TBST containing 0.1% BSA.
  • Washing with TBST for 15min at RT.
  • Incubation with anti-rabbit HRP-linked secondary antibody (Cell Signaling, 7074S, lot 26) diluted 1:2000 in TBST a shaker for 1h at RT.
  • Three rinses, followed by four 15min washes, twice TBST and twice with TBS.
  • Chemiluminescence detection was done using ECL Select Western blotting detection reagent (Amersham, RPN2235, lot 9723881).
  • Blot was developed on a Biorad imaging system.
Experimental Notes
  • The antibody did not consistently pick up the WT mice on normal and low iron diet.
  • Also, there is still a faint band in the NCOA4 -/- lane. We are confident that it is a true KO mouse, as there is high ferritin expression even after 7 days of low iron diet (compared to ferritin expression in WT low iron mice as well as that of the HIF over expression mouse, which are known to have high intestinal ftn content).
Validation Images
Western Blotting validation image for anti-Nuclear Receptor Coactivator 4 (NCOA4) (N-Term) antibody (ABIN2780237) Mouse duodenum whole cell lysates from intestinal HIF overexpression (HIF OE) mice, N...