Recombinant Anti-BTF3 antibody [EPR16495] (ab203517)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16495] to BTF3
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-BTF3 antibody [EPR16495]
See all BTF3 primary antibodies -
Description
Rabbit monoclonal [EPR16495] to BTF3 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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 (intra): HeLa cells. IP: NIH/3T3 whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16495 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
1/250.
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WB | (1) |
1/1000. Detects a band of approximately 22, 18 kDa (predicted molecular weight: 22 kDa).
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ICC/IF |
1/250.
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IP |
1/70.
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IHC-P |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/250. |
WB
1/1000. Detects a band of approximately 22, 18 kDa (predicted molecular weight: 22 kDa). |
ICC/IF
1/250. |
IP
1/70. |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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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
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 689 Human
- Entrez Gene: 218490 Mouse
- Entrez Gene: 294680 Rat
- Omim: 602542 Human
- SwissProt: P20290 Human
- SwissProt: Q64152 Mouse
- Unigene: 591768 Human
- Unigene: 371688 Mouse
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Alternative names
- Basic transcription factor 3 antibody
- BETA NAC antibody
- BTF 3 antibody
see all
Images
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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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 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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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.
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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.
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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.
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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.
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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. -
Intracellular 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.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab203517 has been referenced in 3 publications.
- Zhang Y et al. BTF3 confers oncogenic activity in prostate cancer through transcriptional upregulation of Replication Factor C. Cell Death Dis 12:12 (2021). PubMed: 33414468
- Gordon DE et al. A Quantitative Genetic Interaction Map of HIV Infection. Mol Cell 78:197-209.e7 (2020). PubMed: 32084337
- Wu X et al. miR-802 inhibits the epithelial-mesenchymal transition, migration and invasion of cervical cancer by regulating BTF3. Mol Med Rep 22:1883-1891 (2020). PubMed: 32582971