Smad1/5 抗体 (AA 361-465)
SMAD1/5
适用: 人
IF (p), IF (cc), FACS, IHC (fro), IHC (p), ELISA, ICC
宿主: 兔
Polyclonal
unconjugated
(2 references)
(1 validation)-
- 抗原 See all Smad1/5 (SMAD1/5) products
- Smad1/5 (SMAD1/5) (Smad1/5 Protein (SMAD1/5))
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抗原表位
- AA 361-465
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适用
- 人
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宿主
- 兔
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克隆类型
- 多克隆
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标记
- This Smad1/5 antibody is un-conjugated
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应用范围
- Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), ELISA, Immunocytochemistry (ICC)
- 交叉反应
- 人
- 预测反应
- Mouse,Rat,Dog,Cow,Pig,Horse,Rabbit
- 纯化方法
- Purified by Protein A.
- 免疫原
- KLH conjugated synthetic peptide derived from human Smad 5
- 亚型
- IgG
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anti-Smad1/5 Protein (SMAD1/5) (pSer463), (pSer465) antibody
SMAD1/5 适用: 小鼠, 大鼠 IF (p), IF (cc), WB, IHC (fro), IHC (p), ELISA, ICC 宿主: 兔 Polyclonal unconjugated
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- 应用备注
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ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200
ICC 1:100-500 - 限制
- 仅限研究用
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- by
- Alamo Laboratories Inc
- No.
- #029752
- 日期
- 2014.07.03
- 抗原
- Lot Number
- 120918
- Method validated
- Western Blotting
- Positive Control
- Hela cells
- Negative Control
- c6/36 cells (non-reactive species)
- Notes
- A single major band was observed in the positive control at the correct molecular weight. Several additional faint bands appeared in both the positive and negative controls, but they did not indicate significant non-specific interaction.
- Primary Antibody
- Antigen: SMAD, Mothers Against DPP Homolog 1 (SMAD1) (1:200 dilution)
- Catalog number: ABIN732158
- Lot number: 120918
- Secondary Antibody
- Antibody: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate (1:20,000 dilution)
- Lot number: L170-6515
- Full Protocol
- 1. Total protein extracts were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5 min prior to loading.
- 2. 58 μg of boiled extracts were loaded and resolved on 8-16% SDS-polyacrylamide gel.
- 3. The Thermo Scientific - Spectra Multicolor Broad Range (Cat # 26634) were used as molecular mass markers.
- 4. Proteins were then transferred onto PVDF membrane by wet transfer and protein transfer was confirmed with Ponceau-S staining.
- 5. The PVDF membrane was incubated with 25 mL of blocking buffer [Tris Buffered Saline, pH 7.4 plus 0.1% TW20 (TBST)] containing 5% (W/V) BSA at room temperature for 1 h.
- 6. The membrane was rinsed with TBST once.
- 7. The membrane was immersed with the protein side up in the primary antibody solution (anti-SMAD1; 1:200) in TBST containing 5% (W/V) BSA and incubated for 16 h at 4°C.
- 8. The membrane was rinsed in TBST thrice for 5 min each.
- 9. The membrane was incubated in the HRP-conjugated secondary antibody solution (Goat anti-rabbit IgG-HRP; 1:20,000) in TBST containing 5% (W/V) BSA and incubated for 1 h at room temperature (~26°C) with gentle agitation.
- 10. The membrane was rinsed in TBST thrice for 5 min each.
- 11. The membrane was rinsed in TBS twice for 30 s each.
- 12. Signals were detected with ECL-2 Substrate. The blot was scanned for 300 s.
- 13. The membrane was rinsed three times TBST.
- 14. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 min each.
- 15. The membrane was washed in TBST 2 times for 10 min each.
- 16. Repeated Steps 5-12 with the loading control antibody (anti-Actin; 1:6,000) and its matching secondary antibody (Goat anti-rabbit IgG-HRP; 1:20,000).
- Experimental Notes
- - No experimental challenges noted.
生效 #029752 (Western Blotting)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Figure 1. Western blot of lysates from HeLa cells (Lane 1) and c6/36 cells (Lane 2) probed with anti-SMAD1 (upper panel) or with anti-Actin for loading control (lower panel).]()
Figure 1. Western blot of lysates from HeLa cells (Lane 1) and c6/36 cells (Lane 2) probed with anti-SMAD1 (upper panel) or with anti-Actin for loading control (lower panel).
Full Methods -
- 状态
- Liquid
- 浓度
- 1 μg/μL
- 缓冲液
- 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.
- 储存液
- ProClin
- 注意事项
- This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
- 储存条件
- 4 °C,-20 °C
- 储存方法
- Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
- 有效期
- 12 months
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: "MiR-199a-3p suppresses proliferation and invasion of prostate cancer cells by targeting Smad1." in: Oncotarget, Vol. 8, Issue 32, pp. 52465-52473, (2017) (PubMed).
: "Smurf1 inhibits integrin activation by controlling Kindlin-2 ubiquitination and degradation." in: The Journal of cell biology, Vol. 216, Issue 5, pp. 1455-1471, (2017) (PubMed).
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: "MiR-199a-3p suppresses proliferation and invasion of prostate cancer cells by targeting Smad1." in: Oncotarget, Vol. 8, Issue 32, pp. 52465-52473, (2017) (PubMed).
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- 抗原
- Smad1/5 (SMAD1/5) (Smad1/5 Protein (SMAD1/5))
- 别名
- Smad1 + Smad5 (SMAD1/5 产品)
- 背景
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Synonyms: Mothers against decapentaplegic homolog 5, SMAD5, Mothers against decapentaplegic homolog 1, SMAD1
Background: Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD5 is a receptor-regulated SMAD (R-SMAD). Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD1 is a receptor-regulated SMAD (R-SMAD). SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1. May act synergistically with SMAD4 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression.
- 基因ID
- 4090, 4086
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