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

Details for Product anti-BCLAF1 Antibody No. ABIN151225, Anbieter: Anmelden zum Anzeigen
Antigen
  • 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
Epitop
AA 150-200
14
9
9
7
5
3
3
2
2
1
1
1
1
Reaktivität
Human, Maus
94
33
25
10
4
4
4
3
3
2
2
2
2
2
1
1
1
Wirt
Kaninchen
75
20
Klonalität
Polyklonal
Konjugat
Dieser BCLAF1 Antikörper ist unkonjugiert
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
Applikation
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Western Blotting (WB)
77
40
37
20
16
13
9
4
1
1
Optionen
Hersteller
Anmelden zum Anzeigen
Hersteller Produkt- Nr.
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'Independent Validation' Siegel
Antigen BCLAF1
Validierte Anwendung Western Blotting
Positivkontrolle etoposide treated human DoHH2 cells
Negativkontrolle untreated human DoHH2 cells
Bewertung

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

Primärantikörper ABIN151225
Sekundärantikörper Donkey IgG anti-rabbit IgG peroxidase conjugated (Dianova, 711-035-152)
Protokoll
  • 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).
Anmerkungen
  • 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.

Validierungsbilder
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...

Immunogen 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).
Isotyp IgG
Reinigung Immunogen affinity purified
Andere Bezeichnung BTF (BCLAF1 Antibody Abstract)
Hintergrund Gene Symbol: BCLAF1
Gen-ID 9774
NCBI Accession NP_055554
Forschungsgebiet Transcription Factors, Chromatin and Nuclear Signaling, Apoptosis/Necrosis
Pathways Positive Regulation of Response to DNA Damage Stimulus
Applikationshinweise 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
Kommentare

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

Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 0.2 mg/mL
Buffer TBS and 0.1 % BSA
Buffer contains: 0.09 % Sodium Azide
Konservierungsmittel Sodium azide
Vorsichtsmaßnahmen This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung Do not freeze.
Lagerung 4 °C
Informationen zur Lagerung Store at 4°C. Do not freeze.
Bilder des Herstellers
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...
Produkt verwendet in: 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).

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