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CHAF1B Recombinant Monoclonal Antibody

Datasheet
Code CSB-RA699720A0HU
Size US$210
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Image
  • Western Blot
    Positive WB detected in: COLO205 whole cell lysate(30µg), U937 whole cell lysate, HL-60 whole cell lysate(30µg), HEK293 whole cell lysate(30µg), HeLa whole cell lysate(30µg), HepG2 whole cell lysate(30µg)
    All lanes: CAF1 antibody at 1:1000
    Secondary
    Goat polyclonal to rabbit IgG at 1/50000 dilution
    Predicted band size: 61 kDa
    Observed band size: 70 kDa
    Exposure time: 2min
  • IHC image of CSB-RA699720A0HU diluted at 1:100 and staining in paraffin-embedded human colorectal cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB.
  • IHC image of CSB-RA699720A0HU diluted at 1:100 and staining in paraffin-embedded human tonsil tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB.
  • Immunofluorescence staining of Hela cell with CSB-RA699720A0HU at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).
  • Overlay Peak curve showing MCF-7 cells stained with CSB-RA699720A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*106cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/1*106cells) used under the same conditions. Acquisition of >10,000 events was performed.
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Product Details

Uniprot No.
Q13112
Target Names
CHAF1B
Alternative Names
CAF 1 subunit B antibody; CAF 1A antibody; CAF I 60 kDa subunit antibody; CAF IP60 antibody; CAF-1 antibody; CAF-1 subunit B antibody; CAF-I 60 kDa subunit antibody; CAF-I p60 antibody; CAF1 antibody; CAF1A antibody; CAF1B_HUMAN antibody; CAF1P60 antibody; CHAF 1B antibody; CHAF1B antibody; Chromatin assembly factor 1 subunit B antibody; Chromatin assembly factor I antibody; Chromatin assembly factor I p60 subunit antibody; Human chromatin assembly factor-I p60 subunit antibody; M phase phosphoprotein 7 antibody; M-phase phosphoprotein 7 antibody; MPHOSPH7 antibody; MPP 7 antibody; MPP7 antibody; p60 subunit antibody
Species Reactivity
Human
Immunogen
A synthesized peptide derived from human CHAF1B
Immunogen Species
Homo sapiens (Human)
Conjugate
Non-conjugated
Clonality
Monoclonal
Isotype
Rabbit IgG
Clone No.
5G7
Purification Method
Affinity-chromatography
Concentration
It differs from different batches. Please contact us to confirm it.
Buffer
Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.
Form
Liquid
Tested Applications
ELISA, WB, IHC, IF, FC
Recommended Dilution
Application Recommended Dilution
WB 1:500-1:2000
IHC 1:50-1:200
IF 1:50-1:200
FC 1:50-1:200
Protocols
ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Immunofluorescence (IF) Protocol
Flow Cytometry (FC) Protocol
Troubleshooting and FAQs
Antibody FAQs
Storage
Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
Lead Time
Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Description

CHAF1B, also known as chromatin assembly factor 1 subunit B or p60, serves as an essential component of the CAF-1 complex responsible for depositing histones onto newly synthesized DNA during replication. This histone chaperone plays a critical role in maintaining chromatin structure and epigenetic inheritance, making it a key target for researchers investigating DNA replication, cell cycle regulation, and chromatin dynamics. As an M-phase phosphoprotein, CHAF1B is also relevant to studies examining mitotic progression and cancer cell proliferation.

This recombinant monoclonal antibody, clone 5G7, offers the consistency and reproducibility that demanding experimental workflows require. Generated against a synthetic peptide derived from human CHAF1B and produced using recombinant technology, this antibody provides sequence-defined specificity with minimal lot-to-lot variation, ensuring reliable results across extended research projects.

Validation across multiple applications demonstrates considerable experimental flexibility. Western blot analysis detects CHAF1B in diverse human cell lines including COLO205, U937, HL-60, HEK293, HeLa, and HepG2, with an observed band at approximately 70 kDa. This migration above the predicted 61 kDa molecular weight likely reflects post-translational modifications such as phosphorylation, consistent with CHAF1B's known regulation during the cell cycle. Immunohistochemistry staining has been validated in human colorectal cancer and tonsil tissue sections, while immunofluorescence studies in HeLa cells reveal the expected nuclear localization pattern. Flow cytometry analysis in MCF-7 cells further confirms utility for single-cell applications.

This antibody supports research into chromatin biology, DNA replication mechanisms, and cancer-associated epigenetic alterations, providing a reliable tool for investigating CHAF1B function across multiple experimental platforms.

Usage
For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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Target Background

Function
Complex that is thought to mediate chromatin assembly in DNA replication and DNA repair. Assembles histone octamers onto replicating DNA in vitro. CAF-1 performs the first step of the nucleosome assembly process, bringing newly synthesized histones H3 and H4 to replicating DNA; histones H2A/H2B can bind to this chromatin precursor subsequent to DNA replication to complete the histone octamer.
Gene References into Functions
  1. The volumes of the tumors were similar between those injected with the empty vector and control, but were significantly smaller in the knockdown models, suggesting that the knockdown of the CHAF1B gene inhibited tumor growth. H&E staining revealed that tumors were developed in mice in all groups. PMID: 29767268
  2. All the evaluated malignant tumors showed CAF-1 p60 overexpression, confirming the emerging role of CAF-1 p60 as a new proliferation and prognostic marker for cancer. PMID: 23109837
  3. The levels of the EDNRB, HJURP and p60/CAF-1 proteins were strongly associated with overall survival in high-grade gliomas patients (p<0.001, p<0.001 and p=0.002, respectively), whereas the one of PDLI4 was not (P=0.11). PMID: 24039914
  4. Overexpression of CAF-1/p60, on histological and/or cytological samples, characterizes malignant salivary gland tumours with aggressive behaviour PMID: 21109952
  5. CAF-1 is a proliferation marker in various malignant tumours with prognostic value in renal, endometrial and cervical carcinomas, which supports the value of CAF-1 as a clinical marker of cancer progression. PMID: 21083601
  6. Chromatin Assembly Factor-1/p60 may have a role in progression and metastasis of melanoma PMID: 20178651
  7. model is proposed in which the synergism between hAsf1 and CAF-1 for nucleosome formation during DNA repair is achieved through a transient physical interaction allowing histone delivery from Asf1 to CAF-1 [asf1(anti-silencing function 1)] PMID: 11897662
  8. These data identify chromatin assembly factor 1 as an essential factor not only for S-phase-specific chromatin assembly but also for proliferating cell viability. PMID: 15024074
  9. Downregulated protein level of CAF-1 p60 in adult brain of patients with Down syndrome (DS) in contrast to Alzheimer's disease indicates that it can be considered specific for changes in DS. PMID: 15068244
  10. The N- & C-terminal regions of ASF1a and ASF1b determine the different affinities of these two proteins for HIRA, by contacting regions outside the HIRA B domain. CAF-1 p60 also uses B domain-like motifs for binding to ASF1a, thereby competing with HIRA. PMID: 16980972
  11. overexpression of CAF-1/p60 characterized prostatic cancers with a worse prognosis PMID: 19309489
  12. The histone H3K9 methyltransferase SetDB1 associates with the specific heterochromatin protein 1alpha (HP1alpha)-chromatin assembly factor 1 (CAF1) chaperone complex. PMID: 19498464

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Subcellular Location
Nucleus. Cytoplasm. Note=DNA replication foci. Cytoplasmic in M phase.
Protein Families
WD repeat HIR1 family
Database Links

HGNC: 1911

OMIM: 601245

KEGG: hsa:8208

STRING: 9606.ENSP00000315700

UniGene: Hs.75238

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