WB result of CHAF1b Rabbit mAb
Primary antibody: CHAF1b Rabbit mAb at 1/1000 dilution
Lane 1: Jurkat whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: Caco-2 whole cell lysate 20 µg
Lane 4: 293T whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 61 kDa
Observed MW: 61 kDa
Product Details
Product Details
Product Specification
Host | Rabbit |
Antigen | CHAF1b |
Synonyms | Chromatin assembly factor 1 subunit B, CAF-1 subunit B, Chromatin assembly factor I p60 subunit (CAF-I 60 kDa subunit; CAF-I p60); M-phase phosphoprotein 7, CAF1A, CAF1P60, MPHOSPH7, MPP7 |
Location | Cytoplasm, Nucleus |
Accession | Q13112 |
Clone Number | S-R316 |
Antibody Type | Recombinant mAb |
Isotype | IgG |
Application | WB, IHC-P, ICC, ICFCM, IP |
Reactivity | Hu |
Purification | Protein A |
Concentration | 0.5 mg/ml |
Conjugation | Unconjugated |
Physical Appearance | Liquid |
Storage Buffer | PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300 |
Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
application | dilution | species |
WB | 1:1000 | null |
IHC-P | 1:500 | null |
ICC | 1:500 | null |
ICFCM | 1:500 | null |
IP | 1:50 | null |
Background
Chromatin assembly factor I (CAF-1) is required for the assembly of histone octamers onto newly-replicated DNA. CAF-I is composed of three protein subunits, p50, p60, and p150. CHAF1b corresponds to the p60 subunit and is required for chromatin assembly after replication. It is differentially phosphorylated in a cell cycle-dependent manner. In addition, it is normally found in the nucleus except during mitosis, when it is released into the cytoplasm. This protein is a member of the WD-repeat HIR1 family and may also be involved in DNA repair.
Picture
Picture
Western Blot
FC

Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling CHAF1b antibody at 1/500 dilution (0.1 μg)/ (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
IP

CHAF1b Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating CHAF1b in 0.4 mg Jurkat whole cell lysate.
Western blot was performed on the immunoprecipitate using CHAF1b Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1: Jurkat whole cell lysate 20 µg (Input)
Lane 2: CHAF1b Rabbit mAb IP in Jurkat whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in Jurkat whole cell lysate
Predicted MW: 61 kDa
Observed MW: 61 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human colon. Anti-CHAF1b antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human testis. Anti-CHAF1b antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human tonsil. Anti-CHAF1b antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-CHAF1b antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung adenocarcinoma. Anti-CHAF1b antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human stomach cancer. Anti-CHAF1b antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung squamous cell carcinoma. Anti-CHAF1b antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti-CHAF1b antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).
