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CAF1p55/VPRBP Rabbit pAb

Blank control (Black line): Molt4 (Black).
Primary Antibody (green line): Rabbit Anti-CAF1p55_VPRBP antibody (AP59242)
Dilution: 1 µg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
Dilution: 1 µg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (CAF1p55/VPRBP) Polyclonal Antibody, Unconjugated (AP59242) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Blank control (Black line): Molt4 (Black).
Primary Antibody (green line): Rabbit Anti-CAF1p55_VPRBP antibody (AP59242)
Dilution: 1 µg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
Dilution: 1 µg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	IHC-P, IHC-F, IF
Primary Accession	Q9Y4B6
Reactivity	Rat, Human
Predicted	Mouse, Rabbit
Host	Rabbit
Clonality	Polyclonal
Calculated MW	169007 Da
Physical State	Liquid
Immunogen	KLH conjugated synthetic peptide derived from human p55/DCAF1
Epitope Specificity	221-320/1507
Isotype	IgG
Purity	affinity purified by Protein A
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SUBCELLULAR LOCATION	Cytoplasm. Nucleus.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	P55 is an extensively palmitoylated erythrocyte membrane protein, and a member of the MAGUK family. P55 also resists salt extraction, resulting in a high affinity for the plasma membrane. P55 contains a PDZ/DHR domain, a conserved SH-3 domain that appears to suppress tyrosine kinase activity of various oncoproteins, a 39-amino acid motif that binds to cytoskeletal protein 4.1R, and a guanylate kinase-like domain. Interaction with glycophorin C (GPC) and 4.1R suggests that p55 may play a role in the dynamic regulation in the erythrocyte membrane. In addition, p55 gene expression in vivo may be associated with a CpG island. P55 is constitutively expressed in K562 erythroleukemia cells during erythropoiesis and undergoes a 2-fold amplification after induction.

Additional Information
Gene ID	9730
Other Names	DDB1- and CUL4-associated factor 1 {ECO:0000312\|HGNC:HGNC:30911}, HIV-1 Vpr-binding protein, VprBP, Serine/threonine-protein kinase VPRBP, 2.7.11.1, Vpr-interacting protein, DCAF1 (HGNC:30911), KIAA0800, RIP, VPRBP
Dilution	IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1ug/Test
Storage	Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	DCAF1 (HGNC:30911)
Synonyms	KIAA0800, RIP, VPRBP
Function	Acts both as a substrate recognition component of E3 ubiquitin-protein ligase complexes and as an atypical serine/threonine- protein kinase, playing key roles in various processes such as cell cycle, telomerase regulation and histone modification. Probable substrate-specific adapter of a DCX (DDB1-CUL4-X-box) E3 ubiquitin- protein ligase complex, named CUL4A-RBX1-DDB1-DCAF1/VPRBP complex, which mediates ubiquitination and proteasome-dependent degradation of proteins such as NF2 (PubMed:23063525). Involved in the turnover of methylated proteins: recognizes and binds methylated proteins via its chromo domain, leading to ubiquitination of target proteins by the RBX1-DDB1-DCAF1/VPRBP complex (PubMed:23063525). The CUL4A-RBX1-DDB1- DCAF1/VPRBP complex is also involved in B-cell development: DCAF1 is recruited by RAG1 to ubiquitinate proteins, leading to limit error- prone repair during V(D)J recombination (By similarity). Also part of the EDVP complex, an E3 ligase complex that mediates ubiquitination of proteins such as TERT, leading to TERT degradation and telomerase inhibition (PubMed:19287380, PubMed:23362280). The EDVP complex also mediates ubiquitination and degradation of CCP110 (PubMed:28242748, PubMed:34259627). Also acts as an atypical serine/threonine-protein kinase that specifically mediates phosphorylation of 'Thr-120' of histone H2A (H2AT120ph) in a nucleosomal context, thereby repressing transcription (PubMed:24140421). H2AT120ph is present in the regulatory region of many tumor suppresor genes, down-regulates their transcription and is present at high level in a number of tumors (PubMed:24140421). Involved in JNK-mediated apoptosis during cell competition process via its interaction with LLGL1 and LLGL2 (PubMed:20644714). By acting on TET dioxygenses, essential for oocyte maintenance at the primordial follicle stage, hence essential for female fertility (By similarity).
Cellular Location	Cytoplasm. Nucleus. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. Note=Associated with chromatin in a DDB1- independent and cell cycle-dependent manner: recruited to chromatin as DNA is being replicated and is released from chromatin before mitosis (By similarity). Homogeneous pancellular distribution is observed outside S-phase and a slight cytoplasmic-to-nuclear translocation from early to late S-phase (By similarity). Colocalizes with TET1 and PCNA at replicating heterochromatin during late S phase (By similarity) More concentrated in nuclei than in cytoplasm in germinal vesicle (GV) stage oocytes, zygotes and the 2-cell stage, but distributed in the cytoplasm at the MII-stage oocytes (By similarity). Localizes to centrosomes following interaction with CEP78 (PubMed:28242748, PubMed:34259627). {ECO:0000250\|UniProtKB:Q80TR8, ECO:0000269\|PubMed:28242748, ECO:0000269\|PubMed:34259627}
Tissue Location	Ubiquitously expressed.