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

Details for Product anti-BCLAF1 Antibody No. ABIN151225, Fournisseur: Connectez-vous pour afficher
Antigène
  • 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
Épitope
AA 150-200
14
9
9
7
3
3
3
2
2
2
1
1
1
1
Reactivité
Humain, Souris
96
33
25
10
4
4
4
3
3
2
2
2
2
2
1
1
1
Hôte
Lapin
77
20
Clonalité
Polyclonal
Conjugué
Cet anticorp BCLAF1 est non-conjugé
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
Application
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Western Blotting (WB)
77
40
37
22
16
13
11
4
1
1
Options
Fournisseur
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Supplier Product No.
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'Independent Validation' signe
Antigène BCLAF1
Application validée Western Blotting
Contrôle positif etoposide treated human DoHH2 cells
Contrôle négative untreated human DoHH2 cells
Conclusion

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

Anticorps primaire ABIN151225
Anticorps secondaire Donkey IgG anti-rabbit IgG peroxidase conjugated (Dianova, 711-035-152)
Protocole
  • 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).
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...

Immunogène 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).
Isotype IgG
Purification Immunogen affinity purified
Autre désignation BTF (BCLAF1 Antibody Extrait)
Sujet Gene Symbol: BCLAF1
ID gène 9774
NCBI Accession NP_055554
Domaine de recherche Transcription Factors, Chromatin and Nuclear Signaling, Apoptosis/Necrosis
Pathways Positive Regulation of Response to DNA Damage Stimulus
Indications d'application 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
Commentaires

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

Restrictions For Research Use only
Format Liquid
Concentration 0.2 mg/mL
Buffer TBS and 0.1 % BSA
Buffer contains: 0.09 % Sodium Azide
Agent conservateur Sodium azide
Précaution d'utilisation This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Conseil sur la manipulation Do not freeze.
Stock 4 °C
Stockage commentaire Store at 4°C. Do not freeze.
Supplier Images
Western Blotting (WB) image for anti-BCL2-Associated Transcription Factor 1 (BCLAF1) (AA 150-200) antibody (ABIN151225) Detection of Human BTF by Western Blot and Immunoprecipitation. Samples: Whole cell l...
Immunoprecipitation (IP) image for anti-BCL2-Associated Transcription Factor 1 (BCLAF1) (AA 150-200) antibody (ABIN151225) Immunoprecipitation: BTF Antibody [ABIN151225] - Whole cell lysate from HeLa (5, 15 a...
Western Blotting (WB) image for anti-BCL2-Associated Transcription Factor 1 (BCLAF1) (AA 150-200) antibody (ABIN151225) Western Blot: BTF Antibody [ABIN151225] - : Whole cell lysate (50 ug) from TCMK-1, 4T...
Western Blotting (WB) image for anti-BCL2-Associated Transcription Factor 1 (BCLAF1) (AA 150-200) antibody (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-BCL2-Associated Transcription Factor 1 (BCLAF1) (AA 150-200) antibody (ABIN151225) Immunohistochemistry-Paraffin: BTF Antibody [ABIN151225] - Human skin carcinoma. Anti...
Produit citée dans: Lee, Kalejta, Kerry, Semmes, OConnor, Khan, Garcia, Shenk, Murphy: "BclAF1 restriction factor is neutralized by proteasomal degradation and microRNA repression during human cytomegalovirus infection." dans: 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." dans: 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." dans: 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." dans: Molecular and cellular biology, Vol. 27, Issue 24, pp. 8480-91, 2007 (PubMed).

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