Overview

  • Product name

    Anti-BTF3 antibody [EPR16495]
    See all BTF3 primary antibodies
  • Description

    Rabbit monoclonal [EPR16495] to BTF3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human BTF3 aa 1-150. The exact sequence is proprietary.
    Database link: P20290

  • Positive control

    • WB: MCF7, SK-BR-3, HeLa, Jurkat, C6, RAW 264.7, PC-12 and NIH/3T3 cell lysates; Human lymph node and fetal kidney lysates; Mouse heart and spleen lysates. IHC-P: Human cervix carcinoma, Human spleen, mouse liver and rat testis tissues. ICC/IF: HeLa and SK-BR-3 cells. Flow Cyt: HeLa cells. IP: NIH/3T3 whole cell lysate.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab203517 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 22, 18 kDa (predicted molecular weight: 22 kDa).
ICC/IF 1/250.
IP 1/70.
Flow Cyt 1/250.
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    General transcription factor. BTF3 can form a stable complex with RNA polymerase II. Required for the initiation of transcription.
  • Sequence similarities

    Belongs to the NAC-beta family.
    Contains 1 NAC-A/B (NAC-alpha/beta) domain.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Basic transcription factor 3 antibody
    • BETA NAC antibody
    • BTF 3 antibody
    • btf3 antibody
    • BTF3_HUMAN antibody
    • BTF3a antibody
    • BTF3b antibody
    • NACB antibody
    • Nascent polypeptide associated complex beta polypeptide antibody
    • RNA polymerase B transcription factor 3 antibody
    • Transcription factor BTF3 antibody
    see all

Images

  • All lanes : Anti-BTF3 antibody [EPR16495] (ab203517) at 1/2000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
    Lane 2 : SK-BR-3 (Human mammary gland adenocarcinoma cell line) cell lysate
    Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 18,22 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-BTF3 antibody [EPR16495] (ab203517) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 22 kDa
    Observed band size: 18,22 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-BTF3 antibody [EPR16495] (ab203517) at 1/1000 dilution + C6 (Rat glial tumor cells) cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 18,22 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-BTF3 antibody [EPR16495] (ab203517) at 1/1000 dilution + Human lymph node lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 18 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-BTF3 antibody [EPR16495] (ab203517) at 1/1000 dilution

    Lane 1 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) cell lysate
    Lane 3 : NIH/3T3 (Mouse embyro fibroblast cells) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 18 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute
  • All lanes : Anti-BTF3 antibody [EPR16495] (ab203517) at 1/1000 dilution

    Lane 1 : Mouse heart lysate
    Lane 2 : Mouse spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 22 kDa
    Observed band size: 18 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma labeling BTF3 with ab203517 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on Human cervix carcinoma tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Human spleen labeling BTF3 with ab203517 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on Human spleen tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling BTF3 with ab203517 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on mouse liver tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling BTF3 with ab203517 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on rat testis tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling BTF3 with ab203517 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal imaging showing cytoplasmic and weakly nuclear staining on HeLa cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab203517 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-BR-3 (Human mammary gland adenocarcinoma cell line) cells labeling BTF3 with ab203517 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal imaging showing cytoplasmic and weakly nuclear staining on SK-BR-3 cell line. 

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab203517 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling BTF3 with ab203517 at 1/250 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • BTF3 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with ab203517 at 1/70 dilution.

    Western blot was performed from the immunoprecipitate using ab203517 at 1/1000 dilution.

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: NIH/3T3 whole cell lysate 10ug (Input).

    Lane 2: ab203517 IP in NIH/3T3 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab203517 in NIH/3T3 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

References

ab203517 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Gel Running Conditions
Non-reduced Denaturing (11%)
Loading amount
70 µg
Treatment
BTF3 siRNA
Specification
HeLa cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dikla Vardi

Verified customer

Submitted Jun 20 2017

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