Recombinant Anti-DPF2/REQ antibody [EPR9206(B)] (ab134942)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR9206(B)] to DPF2/REQ
- Suitable for: WB, IP, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-DPF2/REQ antibody [EPR9206(B)]
See all DPF2/REQ primary antibodies -
Description
Rabbit monoclonal [EPR9206(B)] to DPF2/REQ -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide corresponding to Human DPF2/REQ.
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Positive control
- IHC-P: Human kidney, colon and breast carcinoma tissue; mouse and rat cerebral cortex tissue. WB: A431, HeLa, Jurkat, PC-12, NIH/3T3 and LNCaP cell lysates. ICC: HeLa cells.
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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. Avoid freeze / thaw cycle. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 5.00 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR9206(B) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab134942 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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WB |
1/1000 - 1/10000. Predicted molecular weight: 44 kDa.
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IP |
1/10 - 1/100.
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ICC/IF |
1/100 - 1/250.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
For antigen retrieval: heat up to 98°C, below boiling, and then let cool for 10-20 min. Use of an HRP/AP polymerized secondary antibody is recommended. |
Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 44 kDa. |
IP
1/10 - 1/100. |
ICC/IF
1/100 - 1/250. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. For antigen retrieval: heat up to 98°C, below boiling, and then let cool for 10-20 min. Use of an HRP/AP polymerized secondary antibody is recommended. |
Target
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Function
May be a transcription factor required for the apoptosis response following survival factor withdrawal from myeloid cells. Might also have a role in the development and maturation of lymphoid cells. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the requiem/DPF family.
Contains 1 C2H2-type zinc finger.
Contains 2 PHD-type zinc fingers. -
Cellular localization
Nucleus. Cytoplasm. 30% nuclear. 70% cytoplasmic. - Information by UniProt
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Database links
- Entrez Gene: 5977 Human
- Entrez Gene: 19708 Mouse
- Entrez Gene: 361711 Rat
- Omim: 601671 Human
- SwissProt: Q92785 Human
- SwissProt: Q61103 Mouse
- Unigene: 13495 Human
- Unigene: 2651 Mouse
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Alternative names
- Apoptosis response zinc finger protein antibody
- BAF45D antibody
- BRG1-associated factor 45D antibody
see all
Images
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All lanes : Anti-DPF2/REQ antibody [EPR9206(B)] (ab134942) at 1/1000 dilution
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : DPF2 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : LNCaP (Human prostate cancer cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab134942 observed at 50 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab134942 was shown to react with DPF2/REQ in wild-type A431 cells in western blot. Loss of signal was observed when DPF2 knockout sample was used. Wild-type and DPF2 knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab134942 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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ab134942 (purified) at 1/100 dilution (2µg) immunoprecipitating DPF2/REQ in HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate.
Lane 1 (input): HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 10ug
Lane 2 (+): ab134942 + HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab134942 in HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysateFor western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling DPF2/REQ with purified ab134942 at 1/200 dilution (8.3μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889, an anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 μg/ml). ab150077, a Goat anti-rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling DPF2/REQ with purified ab134942 at 1/200 dilution (8.3 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9. Tissue was counterstained with hematoxylin. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebral cortex tissue sections labeling DPF2/REQ with purified ab134942 at 1/200 dilution (8.3 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9. Tissue was counterstained with hematoxylin. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebral cortex tissue sections labeling DPF2/REQ with purified ab134942 at 1/200 dilution (8.3 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9. Tissue was counterstained with hematoxylin. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling DPF2/REQ with purified ab134942 at 1/200 dilution (8.3 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9. Tissue was counterstained with hematoxylin. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.
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Anti-DPF2/REQ antibody [EPR9206(B)] (ab134942) at 1/2000 dilution (purified) + NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 44 kDa
Observed band size: 44 kDaBlocking and diluting buffer: 5% NFDM /TBST.
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All lanes : Anti-DPF2/REQ antibody [EPR9206(B)] (ab134942) at 1/1000 dilution (purified)
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : LNCaP (Human prostate cancer cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 44 kDa
Observed band size: 44 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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All lanes : Anti-DPF2/REQ antibody [EPR9206(B)] (ab134942) at 1/1000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : LNCaP cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution
Developed using the ECL technique.
Predicted band size: 44 kDa -
Immunohistochemical analysis of paraffin-embedded, formalin-fixed Human kidney tissue, labelling DPF2/REQ using unpurified ab134942 at a 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (9)
ab134942 has been referenced in 9 publications.
- Li J et al. A Role for SMARCB1 in Synovial Sarcomagenesis Reveals That SS18-SSX Induces Canonical BAF Destruction. Cancer Discov 11:2620-2637 (2021). PubMed: 34078620
- Sunkel BD et al. Evidence of pioneer factor activity of an oncogenic fusion transcription factor. iScience 24:102867 (2021). PubMed: 34386729
- Zhang S et al. mTORC1 Promotes ARID1A Degradation and Oncogenic Chromatin Remodeling in Hepatocellular Carcinoma. Cancer Res 81:5652-5665 (2021). PubMed: 34429326
- Fernando TM et al. Functional characterization of SMARCA4 variants identified by targeted exome-sequencing of 131,668 cancer patients. Nat Commun 11:5551 (2020). PubMed: 33144586
- Pan J et al. The ATPase module of mammalian SWI/SNF family complexes mediates subcomplex identity and catalytic activity-independent genomic targeting. Nat Genet 51:618-626 (2019). PubMed: 30858614
- Pan J et al. Interrogation of Mammalian Protein Complex Structure, Function, and Membership Using Genome-Scale Fitness Screens. Cell Syst 6:555-568.e7 (2018). WB . PubMed: 29778836
- Local A et al. Identification of H3K4me1-associated proteins at mammalian enhancers. Nat Genet 50:73-82 (2018). PubMed: 29255264
- Mashtalir N et al. Modular Organization and Assembly of SWI/SNF Family Chromatin Remodeling Complexes. Cell 175:1272-1288.e20 (2018). PubMed: 30343899
- Cabot B et al. Differential expression of key subunits of SWI/SNF chromatin remodeling complexes in porcine embryos derived in vitro or in vivo. Mol Reprod Dev 84:1238-1249 (2017). ICC/IF . PubMed: 29024220