WB result of IKAROS Recombinant Rabbit mAb
Primary antibody: IKAROS Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: Raji whole cell lysate 20 µg
Lane 3: Ramos whole cell lysate 20 µg
Lane 4: Molt-4 whole cell lysate 20 µg
Negative control: HeLa whole cell lysate
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 58 kDa
Observed MW: 50~70 kDa
Product Details
Product Details
Product Specification
Host | Rabbit |
Antigen | IKAROS |
Synonyms | DNA-binding protein Ikaros; Ikaros family zinc finger protein 1; Lymphoid transcription factor LyF-1; IKZF1; IK1; LYF1; ZNFN1A1 |
Immunogen | Synthetic Peptide |
Location | Nucleus |
Accession | Q13422 |
Clone Number | S-729-146 |
Antibody Type | Recombinant mAb |
Isotype | IgG |
Application | WB, IHC-P, ICC, IP |
Reactivity | Hu |
Positive Sample | Raji, Ramos, Molt-4 |
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 | Hu |
IP | 1:50 | Hu |
IHC-P | 1:1000 | Hu |
ICC | 1:500 | Hu |
Background
Ikaros is a transcription factor that is encoded by the IKZF genes of the Ikaros family zinc finger group. Zinc finger is a small structural motif of protein that allows protein binding to DNA or RNA molecule that is characterized by the coordination of one or more zinc ions (Zn2+) in order to stabilize the fold. Ikaros displays crucial functions in the hematopoietic system and is a known regulator of immune cells development, mainly in early B cells, CD4+ T cells. Its dysfunction has been linked to the development of chronic lymphocytic leukemia. In particular, Ikaros has been found in recent years to be a major tumor suppressor involved in human B-cell acute lymphoblastic leukemia and that it also has a part in the differentiation and function of individual T helper cells.
Picture
Picture
Western Blot
IP
IKAROS Recombinant Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating IKAROS in 0.4 mg MOLT4 whole cell lysate.
Western blot was performed on the immunoprecipitate using IKAROS Recombinant Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: MOLT4 whole cell lysate 40 µg (Input)
Lane 2: IKAROS Recombinant Rabbit mAb IP in MOLT4 whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in MOLT4 whole cell lysate
Predicted MW: 58 kDa
Observed MW: 50~70 kDa
Exposure time: 180 s
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human tonsil. Anti- IKAROS antibody was used at 1/1000 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- IKAROS antibody was used at 1/1000 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 Ramos cells (top panel) and negative staining in HeLa cells (below panel). Anti- IKAROS 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).
