Product nameAnti-BAF53A antibody - ChIP Grade
See all BAF53A primary antibodies
DescriptionRabbit polyclonal to BAF53A - ChIP Grade
SpecificityIn Western blot, ab3882 detects BAF53 and Tip60-TAP in total extracts of Tip60-TAP. Following TEV elution of Tip60-TAP ab3882 only detects BAF53.
Tested applicationsSuitable for: ChIP, ICC/IF, WB, IHC-P, IHC-Frmore details
Species reactivityReacts with: Mouse, Rat, Human
- This antibody gave a positive signal in the following whole cell lysates: NIH 3T3 (Mouse embryonic fibroblast cell line) MEF1 (Mouse embryonic fibroblast cell line) PC12 (Rat adrenal pheochromocytoma cell line) This antibody gave a positive signal in the following tissue lysates: Skeletal Muscle (Mouse) Ovary (Mouse) - normal tissue
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
ChIP Related Products
Our Abpromise guarantee covers the use of ab3882 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|WB||1/1000. Detects a band of approximately 53 kDa.|
|IHC-P||1/35. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
RelevanceThis protein is a family member of actin-related proteins (ARPs), which share significant amino acid sequence identity to conventional actins. Both actins and ARPs have an actin fold, which is an ATP-binding cleft, as a common feature. The ARPs are involved in diverse cellular processes, including vesicular transport, spindle orientation, nuclear migration and chromatin remodeling. This protein is a 53 kDa subunit of the BAF (BRG1/brm-associated factor) complex in mammals, which is functionally related to SWI/SNF complex in S. cerevisiae and Drosophila; the latter is thought to facilitate transcriptional activation of specific genes by antagonizing chromatin-mediated transcriptional repression. Together with beta-actin, it is required for maximal ATPase activity of BRG1, and for the association of the BAF complex with chromatin/matrix. It is required for maximal ATPase activity of SMARCA4/BRG1 and for association of the SMARCA4/BRG1 containing remodelling complex BAF with chromatin/nuclear matrix. It is a component of the NuA4 histone acetyltransferase (HAT) complex which is involved in transcriptional activation of select genes principally by acetylation of nucleosomal histone H4 and H2A. This modification may both alter nucleosome - DNA interactions and promote interaction of the modified histones with other proteins which positively regulate transcription. This complex may be required for the activation of transcriptional programs associated with oncogene and proto-oncogene mediated growth induction, tumor suppressor mediated growth arrest and replicative senescence, apoptosis, and DNA repair. NuA4 may also play a direct role in DNA repair when recruited to sites of DNA damage.
- 53 kDa BRG1 associated factor A antibody
- Actin like 6A antibody
- Actin like protein 6A antibody
All lanes : Anti-BAF53A antibody - ChIP Grade (ab3882) at 1/1000 dilution
Lane 1 : TEV elution of Tip60-TAP clone 1
Lane 2 : TEV elution of Tip60-TAP clone 2
Lane 3 : Total extract of Tip60-TAP clone 1
Lane 4 : Total extract of Tip60-TAP clone 2
Performed under reducing conditions.
Predicted band size: 47 kDa
Total extracts from two different clones of MCF7 cells expressing a TAP tagged version of Tip60 were partially purified over IgG sepharose resin and eluted with TEV protease. Western blot was performed using a 1/1000 dilution of the BAF53A antibody.
Lane 1. TEV elution of Tip60-TAP clone #1
Lane 2. TEV elution of Tip60-TAP clone #2
Lane 3. Total extract of Tip60-TAP clone #1
Lane 4. Total extract of Tip60-TAP clone #2
NB: Tip60 is detected in the total extract due to the presence of Protein A in the TAP Tag (to which the primary and secondary abs bind). The TEV protease removes the Tag in lanes 1 and 2.
Sonicated Chromatin prepared from untreated (UI) or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure with ab3882 to BAF53A and the immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are nomalized over inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 8 µl of ab3882 and 2x106 cells were used in each ChIP experiment.
ab3882 at a 1/200 dilution staining asynchronous HeLa cells by ICC/IF. The cells were paraformaldehyde fixed and incubated with the antibody for 30 minutes at room temperature. Bound antibody was detected using a Cy3 conjugated Goat anti-rabbit antibody. Nuclei were visualised using DAPI stain. ab3882 gives a predominantly nuclear staining pattern.
This image is courtesy of an Abreview submitted by Kirk McManus.
All lanes : Anti-BAF53A antibody - ChIP Grade (ab3882) at 1 µg/ml
Lane 1 :
NIH 3T3 whole cell lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 :
Mouse skeletal muscle tissue lysate - total protein (ab29711)
Lane 4 : Ovary (Mouse) Tissue Lysate - normal tissue
Lane 5 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 47 kDa
Observed band size: 47 kDa
Additional bands at: 120 kDa, 18 kDa. We are unsure as to the identity of these extra bands.
Image courtesy of Human Protein Atlas
ab3882 staining BAF53A in human ovary, showing a distinct and strong staining pattern of the nucleus of the ovarian stromal cells. Paraffin-embedded ovary tissue was incubated with ab3882 (1/35) for 30 minutes at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab3882 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
This product has been referenced in:
- Masuda Y et al. TRIM29 regulates the assembly of DNA repair proteins into damaged chromatin. Nat Commun 6:7299 (2015). Human . Read more (PubMed: 26095369) »
- Lu W et al. Actl6a protects embryonic stem cells from differentiating into primitive endoderm. Stem Cells 33:1782-93 (2015). Read more (PubMed: 25802002) »