NCOA4 抗体 (Nuclear Receptor Coactivator 4) (N-Term)

Details for Product anti-NCOA4 Antibody No. ABIN2780237, 供应商: Log in to see
抗原
  • 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
抗原表位
N-Term
12
10
9
9
8
8
6
4
3
3
3
2
2
1
1
1
1
适用
牛, 犬, 豚鼠, 马, 人, 小鼠, 兔, 大鼠
85
12
9
3
3
3
2
1
1
1
宿主

45
40
克隆类型
多克隆
标记
This NCOA4 antibody is un-conjugated
3
3
3
3
3
3
应用范围
Western Blotting (WB)
75
39
21
9
8
4
2
1
1
选项
Supplier
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'独立验证'标志
抗原 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Notes Passed, based on the fact that the antibody picked up the positive control (unpublished data suggests that NCOA4 is highly expressed in an intestinal HIF overexpression model).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)
实验流程
  • 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...
免疫原 The immunogen is a synthetic peptide corresponding to a region of Mouse
Blocking Peptide Blocking peptide for this product available: ABIN980536
序列 CLIHQLEYTQ NKDLANQVSV CLERLGSLAL KPEDSTVLLF EADTSALRQT
预测反应 Cow: 92%, Dog: 93%, Guinea Pig: 100%, Horse: 93%, Human: 100%, Mouse: 100%, Rabbit: 100%, Rat: 100%
产品特性 This is a rabbit polyclonal antibody against Ncoa4. It was validated on Western Blot.
纯化方法 Affinity Purified
别名 Ncoa4 (NCOA4 Antibody 摘要)
背景 The function of Ncoa4 remains unknown.
Alias Symbols: 1110034E15Rik, 4432406M01Rik, AI227008, ARA70, MGC103382, Rfg
Protein Interaction Partner: Ndn,
Protein Size: 625
分子量 70 kDa
基因ID 27057
NCBI登录号 NM_001033988, NP_001029160
途径 Intracellular Steroid Hormone Receptor Signaling Pathway
应用备注 Optimal working dilutions should be determined experimentally by the investigator.
说明

Antigen size: 625 AA

限制 仅限研究用
状态 Liquid
浓度 Lot specific
缓冲液 Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.
储存液 Sodium azide
注意事项 This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
注意事项 Avoid repeated freeze-thaw cycles.
储存条件 -20 °C
储存方法 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.
厂商提供的图像
Western Blotting (WB) image for anti-Nuclear Receptor Coactivator 4 (NCOA4) (N-Term) antibody (ABIN2780237) anti-Nuclear Receptor Coactivator 4 (NCOA4) (N-Term) antibody
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