Product: IKZF1 Antibody
Catalog: DF6659
Description: Rabbit polyclonal antibody to IKZF1
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Horse, Rabbit, Dog
Mol.Wt.: 50-70kD; 58kD(Calculated).
Uniprot: Q13422
RRID: AB_2838621

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(92%), Horse(92%), Rabbit(85%), Dog(92%)
Clonality:
Polyclonal
Specificity:
IKZF1 Antibody detects endogenous levels of total IKZF1.
RRID:
AB_2838621
Cite Format: Affinity Biosciences Cat# DF6659, RRID:AB_2838621.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

CLL associated antigen KW 6; DNA-binding protein Ikaros; hIk 1; Hs.54452; IK1; Ikaros (zinc finger protein); IKAROS; IKAROS family zinc finger 1 (Ikaros); Ikaros family zinc finger protein 1; Ikzf1; IKZF1_HUMAN; LYF1; Lymphoid transcription factor LyF-1; PRO0758; Zinc finger protein subfamily 1A 1 (Ikaros); Zinc finger protein subfamily 1A 1; Zinc finger protein, subfamily 1A, member 1; ZNFN1A1;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q13422 IKZF1_HUMAN:

Abundantly expressed in thymus, spleen and peripheral blood Leukocytes and lymph nodes. Lower expression in bone marrow and small intestine.

Description:
Ikaros family proteins, characterized by the presence of an N-terminal zinc finger DNA-binding domain and a C-terminal dimerization domain, belong to the Kruppel transcription factor superfamily. The Ikaros family includes Ikaros, Aiolos, Helios, and possibly EOS and Pegasus (1). They can form homodimers and heterodimers with other members of the Ikaros family. Due to differential splicing, multiple isoforms can be generated and some behave in a dominant negative manner upon dimerization (2).Ikaros, also known as Ikaros family zinc finger protein 1 (IKZF1) and Lymphoid transcription factor 1 (LYF-1), is expressed abundantly in blood cells. Genetic studies in mice demonstrate that Ikaros is important for the normal development of B, T, natural killer, and dendritic cells, and that it functions as a tumor suppressor (3,4). Research studies have shown that imbalanced expression of different Ikaros isoforms, as well as deletions of Ikaros are associated with hematologic malignancies in humans (5-7).
Sequence:
MDADEGQDMSQVSGKESPPVSDTPDEGDEPMPIPEDLSTTSGGQQSSKSDRVVASNVKVETQSDEENGRACEMNGEECAEDLRMLDASGEKMNGSHRDQGSSALSGVGGIRLPNGKLKCDICGIICIGPNVLMVHKRSHTGERPFQCNQCGASFTQKGNLLRHIKLHSGEKPFKCHLCNYACRRRDALTGHLRTHSVGKPHKCGYCGRSYKQRSSLEEHKERCHNYLESMGLPGTLYPVIKEETNHSEMAEDLCKIGSERSLVLDRLASNVAKRKSSMPQKFLGDKGLSDTPYDSSASYEKENEMMKSHVMDQAINNAINYLGAESLRPLVQTPPGGSEVVPVISPMYQLHKPLAEGTPRSNHSAQDSAVENLLLLSKAKLVPSEREASPSNSCQDSTDTESNNEEQRSGLIYLTNHIAPHARNGLSLKEEHRAYDLLRAASENSQDALRVVSTSGEQMKVYKCEHCRVLFLDHVMYTIHMGCHGFRDPFECNMCGYHSQDRYEFSSHITRGEHRFHMS

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
92
Horse
92
Dog
92
Rabbit
85
Sheep
77
Xenopus
77
Zebrafish
71
Chicken
62
Bovine
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q13422 As Substrate

Site PTM Type Enzyme
S13 Phosphorylation P67870 (CSNK2B) , P68400 (CSNK2A1)
T23 Phosphorylation P68400 (CSNK2A1) , P67870 (CSNK2B)
K58 Sumoylation
T61 Phosphorylation
S63 Phosphorylation P68400 (CSNK2A1) , P67870 (CSNK2B)
R83 Methylation
S88 Phosphorylation
K91 Ubiquitination
S95 Phosphorylation
S101 Phosphorylation P68400 (CSNK2A1) , P67870 (CSNK2B)
S105 Phosphorylation
K116 Acetylation
T140 Phosphorylation
K157 Ubiquitination
K165 Ubiquitination
S168 Phosphorylation
K171 Acetylation
K171 Sumoylation
K171 Ubiquitination
K174 Ubiquitination
S214 Phosphorylation Q06187 (BTK)
S215 Phosphorylation Q06187 (BTK)
K220 Ubiquitination
K241 Sumoylation
K241 Ubiquitination
K255 Ubiquitination
S258 Phosphorylation
S261 Phosphorylation
S269 Phosphorylation
K273 Ubiquitination
K281 Ubiquitination
K286 Ubiquitination
S289 Phosphorylation
T291 Phosphorylation
Y293 Phosphorylation
S295 Phosphorylation P68400 (CSNK2A1) , P67870 (CSNK2B)
S296 Phosphorylation
S298 Phosphorylation
Y299 Phosphorylation
K301 Ubiquitination
Y321 Phosphorylation
T333 Phosphorylation
S345 Phosphorylation
Y348 Phosphorylation
T358 Phosphorylation
S361 Phosphorylation P43405 (SYK)
S364 Phosphorylation P43405 (SYK)
S368 Phosphorylation
S377 Phosphorylation
K378 Ubiquitination
K380 Ubiquitination
S389 Phosphorylation
S391 Phosphorylation
S393 Phosphorylation
S397 Phosphorylation
T398 Phosphorylation
S402 Phosphorylation
S409 Phosphorylation
Y413 Phosphorylation
T415 Phosphorylation
S427 Phosphorylation
K429 Sumoylation
K429 Ubiquitination
Y435 Phosphorylation
S442 Phosphorylation
S445 Phosphorylation
K460 Ubiquitination

Research Backgrounds

Function:

Transcription regulator of hematopoietic cell differentiation. Binds gamma-satellite DNA. Plays a role in the development of lymphocytes, B- and T-cells. Binds and activates the enhancer (delta-A element) of the CD3-delta gene. Repressor of the TDT (fikzfterminal deoxynucleotidyltransferase) gene during thymocyte differentiation. Regulates transcription through association with both HDAC-dependent and HDAC-independent complexes. Targets the 2 chromatin-remodeling complexes, NuRD and BAF (SWI/SNF), in a single complex (PYR complex), to the beta-globin locus in adult erythrocytes. Increases normal apoptosis in adult erythroid cells. Confers early temporal competence to retinal progenitor cells (RPCs) (By similarity). Function is isoform-specific and is modulated by dominant-negative inactive isoforms.

PTMs:

Phosphorylation controls cell-cycle progression from late G(1) stage to S stage. Hyperphosphorylated during G2/M phase. Dephosphorylated state during late G(1) phase. Phosphorylation on Thr-140 is required for DNA and pericentromeric location during mitosis. CK2 is the main kinase, in vitro. GSK3 and CDK may also contribute to phosphorylation of the C-terminal serine and threonine residues. Phosphorylation on these C-terminal residues reduces the DNA-binding ability. Phosphorylation/dephosphorylation events on Ser-13 and Ser-295 regulate TDT expression during thymocyte differentiation. Dephosphorylation by protein phosphatase 1 regulates stability and pericentromeric heterochromatin location. Phosphorylated in both lymphoid and non-lymphoid tissues (By similarity). Phosphorylation at Ser-361 and Ser-364 downstream of SYK induces nuclear translocation.

Sumoylated. Simulataneous sumoylation on the 2 sites results in a loss of both HDAC-dependent and HDAC-independent repression. Has no effect on pericentromeric heterochromatin location. Desumoylated by SENP1 (By similarity).

Polyubiquitinated.

Subcellular Location:

Nucleus.
Note: In resting lymphocytes, distributed diffusely throughout the nucleus. Localizes to pericentromeric heterochromatin in proliferating cells. This localization requires DNA binding which is regulated by phosphorylation / dephosphorylation events.

Nucleus.
Note: In resting lymphocytes, distributed diffusely throughout the nucleus. Localizes to pericentromeric heterochromatin in proliferating cells. This localization requires DNA binding which is regulated by phosphorylation / dephosphorylation events (By similarity).

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Abundantly expressed in thymus, spleen and peripheral blood Leukocytes and lymph nodes. Lower expression in bone marrow and small intestine.

Subunit Structure:

Heterodimer formed by the various isoforms; this modulates transcription regulator activity. Heterodimer with other IKAROS family members. Interacts with IKZF4 AND IKZF5. Component of the chromatin-remodeling NuRD repressor complex which includes at least HDAC1, HDAC2, RBBP4, RBBP7, IKZF1, MTA2, MBD2, MBD3, MTA1L1, CHD3 and CHD4. Interacts directly with the CHD4 component of the NuRD complex. Interacts directly with SMARCA4; the interaction associates IKFZ1 with the BAF complex. Interacts with SUMO1; the interaction sumoylates IKAROS, promoted by PIAS2 and PIAS3. Interacts with PIAS2 (isoform alpha); the interaction promotes sumoylation and reduces transcription repression. Interacts, to a lesser extent, with PIAS3. Interacts with PPP1CC; the interaction targets PPP1CC to pericentromeric heterochromatin, dephosphorylates IKAROS, stabilizes it and prevents it from degradation. Interacts with IKZF3 (By similarity).

Family&Domains:

The N-terminal zinc-fingers 2 and 3 are required for DNA binding as well as for targeting IKFZ1 to pericentromeric heterochromatin.

The C-terminal zinc-finger domain is required for dimerization.

Belongs to the Ikaros C2H2-type zinc-finger protein family.

References

1). Chidamide-Induced Accumulation of Reactive Oxygen Species Increases Lenalidomide Sensitivity Against Multiple Myeloma Cells. OncoTargets and Therapy, 2021 (PubMed: 34262292) [IF=2.7]

Application: WB    Species: Human    Sample: ARP-1 and RPMI-8226 cells

Figure 5 Chidamide (CHI)-induced ROS accumulation enhances anti-myeloma effect of lenalidomide (Len) through elevating H2O2. (A and B) ARP-1 and RPMI-8226 cells were treated with different doses of lenalidomide (0, 5 and 20 μM) for indicated time (0, 3, 6 and 12 hours), pretreated with or without 15 mmol/L NAC. Then ROS levels were measured using Reactive Oxygen Species Assay Kit. NS, p > 0.05; *, p < 0.05; **, p < 0.01. (C and D) ARP-1 and RPMI-8226 cells were treated with different doses of lenalidomide (0, 5 and 20 μM) for 12 hours, or 1 μM chidamide and 20 μM lenalidomide for 6 hours. Then H2O2 levels were tested using hydrogen peroxide assay kit. (E and F) after treated with 1 μM chidamide, 20 μM lenalidomide or H2O2 (100 and 500 μM) for 6 hours, expression of IKZF1 and IKZF3 were determined using Western blotting in ARP-1 and RPMI-8226 cells. (G and H) pretreated with or without 15 mmol/L NAC, ARP-1 and RPMI-8226 cells were then treated with 1 μM chidamide, 20 μM lenalidomide or 20 μM H2O2 for 24 hours, and CCK-8 assays were used to evaluate the cell viability. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

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