BCLAF1 抗体 (BCL2-Associated Transcription Factor 1) (AA 150-200)

Details for Product anti-BCLAF1 Antibody No. ABIN151225, 供应商: Log in to see
抗原
  • BCLAF1
  • btf
  • bk211l9.1
  • DKFZp468F0113
  • BTF
  • bK211L9.1
  • 2610102K23Rik
  • 2700025J07Rik
  • 2810454G14Rik
  • 5730534O06Rik
  • AI450190
  • AW556225
  • Btf
  • mKIAA0164
  • Aa2-041
  • Fam54a
  • Mtfr2
  • BCL2-associated transcription factor 1
  • BCLAF1
  • bclaf1
  • LOC100231845
  • LOC100339738
  • LOC100351429
  • Bclaf1
Alternatives
anti-小鼠 BCLAF1 抗体 for ELISA
抗原表位
AA 150-200
14
9
9
7
5
3
3
2
1
1
1
1
1
适用
人, 小鼠
93
33
25
10
4
4
4
3
3
2
2
2
2
2
1
1
1
宿主

74
20
克隆类型
多克隆
标记
This BCLAF1 antibody is un-conjugated
4
4
4
3
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
应用范围
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Western Blotting (WB)
76
40
37
20
15
13
9
4
1
1
Supplier
Log in to see
Supplier Product No.
Log in to see

Available images

'独立验证'标志
抗原 BCLAF1
Method validated Western Blotting
Positive Control etoposide treated human DoHH2 cells
Negative Control untreated human DoHH2 cells
Notes

Passed. The BCLAF1 antibody ABIN151225 specifically recognizes BCLAF1 proteins in extracts of etoposide treated human DoHH2 cells.

Primary Antibody ABIN151225
Secondary Antibody Donkey IgG anti-rabbit IgG peroxidase conjugated (Dianova, 711-035-152)
实验流程
  • Grow DoHH2 cells in RPMI Medium 1640 (Gibco, 61870-010, 1870731) supplemented with Fetal Bovine Serum (Gibco, 10270-106, 42Q0271K) at 37°C and 5% CO2.
  • Treat cells for 0 to 8h with 10µM etoposide.
  • Trypsinize cells and wash them with PBS (Gibco, 14190-094, 1852720).
  • Lyse cells in cold IP lysis buffer (20mM Hepes/KOH pH7.4, 110mM K-phosphate, 2mM MgCl2, 150mM NaCl, 0.1% Triton-X, 0.1% Tween-20) containing phosSTOP protease (Roche, 04906837001, lot 12057300) and phosphatase cocktail(cOmplete tablets, Roche, 04693124001, lot 18022500).
  • Quantify total soluble protein in the cell lysates using Roti-Quant protein quantification assay (ROTH, K015.1, lot 0320161148).
  • Dilute 15µg of soluble protein in 10µl 5:1 with 5x Laemmli SDS sample buffer and heat samples up for 5min to 95°C.
  • Separate the samples on a denaturing SDS-PAGE GEL (Expedeon, NXG42012K, lot 151111100) at 120V for 3h.
  • Transfer proteins onto a PVDF membrane (ROTH, HP40.1, lot 486636559) in a Trans-Blot Turbo transfer system (Bio-Rad).
  • Block the membrane with PBS containing 0.1% Tween 20 (PBST) and 2% BSA for 1h at RT.
  • Incubation with primary
    • rabbit anti-BCLAF1 antibody (antibodies-online, ABIN151225) or
    • mouse anti-beta-actin antibody (Santa Cruz Technology, sc-47778, lot F0817)
  • diluted 1:1000 in PBST (PBS containing 0.1% Tween 20) ON at 4°.
  • Wash membrane 3x for 10min with PBST.
  • Incubation with secondary
    • Donkey IgG anti-rabbit IgG peroxidase conjugated (Dianova, 711-035-152)
    • Donkey IgG anti-mouse IgG peroxidase conjugated (Dianova, 715-036-150)
  • diluted 1:5000 in PBST for 2h at RT.
  • Wash membrane 3x for 10min with PBST.
  • Reveal protein bands with Western Bright ECL HRP substrate (Advansta, K-12045, lot 160108-99).
Experimental Notes
  • According to Shao et al. (2016) BCLAF1 implicated in DDR was upregulated during the therapeutic drug doxorubicin-induced senescence. BCLAF1 is a γH2AX-interacting tumor suppressor and is involved in the γH2AX-mediated regulation of apoptosis and DNA repair.

  • Using the BCLAF1 antibody ABIN151225 we got the expected result using cell extracts from etoposide treated human DoHH2 cells. The observed upregulation of BCLAF1 in the lysates correlates with the time the cells were exposed to the drug.

Validation Images
Western Blotting validation image for anti-BCL2-Associated Transcription Factor 1 (BCLAF1) (AA 150-200) antibody (ABIN151225)

Western blot detection of BCLAF1 (upper panel) in lysates of DoHH2 cells treated f...

免疫原 The immunogen recognized by this antibody maps to a region between residues 150 and 200 of human BCL2-Associated Transcription Factor 1 using the numbering given in entry NP_055554.1 (GeneID 9774).
亚型 IgG
纯化方法 Immunogen affinity purified
别名 BTF (BCLAF1 Antibody 摘要)
背景 Gene Symbol: BCLAF1
基因ID 9774
NCBI登录号 NP_055554
研究领域 Transcription Factors, Chromatin and Nuclear Signaling, Apoptosis/Necrosis
途径 Positive Regulation of Response to DNA Damage Stimulus
应用备注 Western Blot 1:2000-1:10000, Immunohistochemistry 1:100 - 1:1000, Immunocytochemistry/Immunofluorescence 1:50 - 1:500, Immunoprecipitation 1 to 4 μg/mg of lysate, Immunohistochemistry-Paraffin 1:10-1:500
说明

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

限制 仅限研究用
状态 Liquid
浓度 0.2 mg/mL
缓冲液 TBS and 0.1 % BSA
Buffer contains: 0.09 % Sodium Azide
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 Do not freeze.
储存条件 4 °C
储存方法 Store at 4°C. Do not freeze.
厂商提供的图像
Western Blotting (WB) image for anti-BCLAF1 抗体 (BCL2-Associated Transcription Factor 1) (AA 150-200) (ABIN151225) Detection of Human BTF by Western Blot and Immunoprecipitation. Samples: Whole cell l...
Immunoprecipitation (IP) image for anti-BCLAF1 抗体 (BCL2-Associated Transcription Factor 1) (AA 150-200) (ABIN151225) Immunoprecipitation: BTF Antibody [ABIN151225] - Whole cell lysate from HeLa (5, 15 a...
Western Blotting (WB) image for anti-BCLAF1 抗体 (BCL2-Associated Transcription Factor 1) (AA 150-200) (ABIN151225) Western Blot: BTF Antibody [ABIN151225] - : Whole cell lysate (50 ug) from TCMK-1, 4T...
Western Blotting (WB) image for anti-BCLAF1 抗体 (BCL2-Associated Transcription Factor 1) (AA 150-200) (ABIN151225) Western Blot: BTF Antibody [ABIN151225] - Whole cell lysate from HeLa (5, 15, 50 ug l...
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) image for anti-BCLAF1 抗体 (BCL2-Associated Transcription Factor 1) (AA 150-200) (ABIN151225) Immunohistochemistry-Paraffin: BTF Antibody [ABIN151225] - Human skin carcinoma. Anti...
有引用在: Lee, Kalejta, Kerry, Semmes, OConnor, Khan, Garcia, Shenk, Murphy: "BclAF1 restriction factor is neutralized by proteasomal degradation and microRNA repression during human cytomegalovirus infection." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 109, Issue 24, pp. 9575-80, 2012 (PubMed).

Lee, Yu, Gunawardena, Xie, Chen: "BCLAF1 is a radiation-induced H2AX-interacting partner involved in γH2AX-mediated regulation of apoptosis and DNA repair." in: Cell death & disease, Vol. 3, pp. e359, 2012 (PubMed).

Ziegelbauer, Sullivan, Ganem: "Tandem array-based expression screens identify host mRNA targets of virus-encoded microRNAs." in: Nature genetics, Vol. 41, Issue 1, pp. 130-4, 2008 (PubMed).

Liu, Lu, Miki, Yoshida: "Protein kinase C delta induces transcription of the TP53 tumor suppressor gene by controlling death-promoting factor Btf in the apoptotic response to DNA damage." in: Molecular and cellular biology, Vol. 27, Issue 24, pp. 8480-91, 2007 (PubMed).