Anti-Ikaros 抗体 [EPR25259-176] (ab300405)

製品名

Anti-Ikaros antibody [EPR25259-176]
Ikaros 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR25259-176] to Ikaros
由来種

Rabbit
特異性

This product could react with Ikaros/IKZF1 isofroms 1-7 theoretically.
アプリケーション

適用あり: ChIP-sequencing, IHC-Fr, Flow Cyt (Intra), IHC-P, IP, WBmore details
適用なし: ChIP or ICC/IF
種交差性

交差種: Mouse, Rat, Human
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- WB: Jurkat, Raji, Daudi, and EL4 whole cell lysates; Mouse and rat spleen and thymus tissue lysates; His-tagged human Ikaros recombinant protein. IHC-P: Human spleen and thymus tissue. Mouse and rat spleen tissue. IHC-FR: Mouse and rat thymus fresh frozen tissue. Flow Cyt (Intra): Jurkat and EL4 cells. IP: Raji and EL4 whole cell lysates.
特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
バッファー

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR25259-176
アイソタイプ

IgG
研究分野
- Cancer
- Cell Death
- Apoptosis
- Other

Alternative Versions
- Anti-Ikaros antibody [EPR25259-176] (BSA and Azide free) (ab300417)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab300405の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
ChIP-sequencing		Use 8µg for 10⁷ cells.
IHC-Fr		1/100. Not Applicable.
Flow Cyt (Intra)		1/500.
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP		1/30.
WB		1/1000. Predicted molecular weight: 57 kDa.

特記事項
ChIP-sequencing Use 8µg for 10⁷ cells.
IHC-Fr 1/100. Not Applicable.
Flow Cyt (Intra) 1/500.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/30.
WB 1/1000. Predicted molecular weight: 57 kDa.

追加情報

Is unsuitable for ChIP or ICC/IF.

機能

Transcription regulator of hematopoietic cell differentiation (PubMed:17934067). Binds gamma-satellite DNA (PubMed:17135265, PubMed:19141594). 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 (PubMed:17135265, PubMed:17934067).
組織特異性

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

Defects in IKZF1 are frequent occurrences (28.6%) in acute lymphoblasic leukemia (ALL). Such alterations or deletions lead to poor prognosis for ALL.
Chromosomal aberrations involving IKZF1 are a cause of B-cell non-Hodgkin lymphomas (B-cell NHL). Translocation t(3;7)(q27;p12), with BCL6.
配列類似性

Belongs to the Ikaros C2H2-type zinc-finger protein family.
Contains 6 C2H2-type zinc fingers.
ドメイン

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.
翻訳後修飾

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.
Polyubiquitinated.
細胞内局在

Cytoplasm; Nucleus. 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 and Nucleus. 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).
Target information above from: UniProt accession Q13422 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 10320 Human
- Entrez Gene: 22778 Mouse
- Entrez Gene: 305501 Rat
- Omim: 603023 Human
- SwissProt: Q13422 Human
- SwissProt: Q03267 Mouse
- Unigene: 435949 Human
- Unigene: 488251 Human
- Unigene: 646004 Human
- Unigene: 731495 Human
- Unigene: 103545 Mouse
see all
製品の状態

There are 7 isoforms produced by alternative splicing.
別名
- CLL associated antigen KW 6 antibody
- DNA-binding protein Ikaros antibody
- hIk 1 antibody
- Hs.54452 antibody
- IK1 antibody
- Ikaros (zinc finger protein) antibody
- IKAROS antibody
- IKAROS family zinc finger 1 (Ikaros) antibody
- Ikaros family zinc finger protein 1 antibody
- Ikzf1 antibody
- IKZF1_HUMAN antibody
- LYF1 antibody
- Lymphoid transcription factor LyF-1 antibody
- PRO0758 antibody
- Zinc finger protein subfamily 1A 1 (Ikaros) antibody
- Zinc finger protein subfamily 1A 1 antibody
- Zinc finger protein, subfamily 1A, member 1 antibody
- ZNFN1A1 antibody
see all

Western blot - Anti-Ikaros antibody [EPR25259-176] (AB300405)

All lanes : Anti-Ikaros antibody [EPR25259-176] (ab300405) at 1/1000 dilution

Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate
Lane 3 : Daudi (human burkitt's lymphoma lymphoblast) whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 5 : EL4 (mouse lymphoma t lymphocyte) whole cell lysate
Lane 6 : Mouse spleen tissue lysate
Lane 7 : Mouse thymus tissue lysate
Lane 8 : Mouse kidney tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 57 kDa
Observed band size: 30-70 kDa why is the actual band size different from the predicted?

Blocking / Diluting buffer: 5% NFDM/TBST.

Exposure time: Lanes 1-4: 59 seconds; Lane 5: 26 seconds; Lanes 6-8: 59 seconds.

This product could react with Ikaros isofroms 1-7 theoretically.

Negative control: HeLa (HPA data); kidney (PMID: 1439790).
Western blot - Anti-Ikaros antibody [EPR25259-176] (AB300405)

All lanes : Anti-Ikaros antibody [EPR25259-176] (ab300405) at 1/1000 dilution

Lane 1 : Rat thymus tissue lysate
Lane 2 : Rat spleen tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 57 kDa
Observed band size: 30-70 kDa why is the actual band size different from the predicted?

Exposure time: 70 seconds

Blocking / Diluting buffer: 5% NFDM/TBST.
Western blot - Anti-Ikaros antibody [EPR25259-176] (AB300405)

All lanes : Anti-Ikaros antibody [EPR25259-176] (ab300405) at 1/1000 dilution

Lane 1 : His-tagged human Ikaros/IKZF1 recombinant protein, 10 ng
Lane 2 : GST-tagged human Helios/IKZF2 recombinant protein, 10 ng
Lane 3 : GST-tagged human Aiolos/IKZF3 recombinant protein, 10 ng

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 57 kDa

Exposure time: 3 seconds

Blocking / Diluting buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Ikaros with ab300405 at 1/2000 dilution (0.27 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Nuclear staining on human spleen. The section was incubated with ab300405 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Immunohistochemical analysis of paraffin-embedded human thymus tissue labeling Ikaros with ab300405 at 1/2000 dilution (0.27 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Nuclear staining on human thymus. The section was incubated with ab300405 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Ikaros with ab300405 at 1/2000 dilution (0.27 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Nuclear staining on mouse spleen. The section was incubated with ab300405 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Ikaros with ab300405 at 1/2000 dilution (0.27 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Nuclear staining on rat spleen. The section was incubated with ab300405 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Frozen sections) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh frozen sectioned mouse thymus tissue labeling Ikaros with ab300405 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (green). Positive staining on mouse thymus is observed. The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

No antigen retrieval was carried out.
Immunohistochemistry (Frozen sections) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh frozen sectioned mouse kidney tissue labeling Ikaros with ab300405 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150081) at 1/1000 dilution (green). The nuclear counter stain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150081) at 1/1000 dilution.

No antigen retrieval was carried out.

Negative control (PMID: 1439790).

No staining on mouse kidney.
Immunohistochemistry (Frozen sections) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh frozen sectioned rat thymus tissue labeling Ikaros with ab300405 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (green). Positive staining on rat thymus is observed. The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

No antigen retrieval was carried out.
Immunohistochemistry (Frozen sections) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh frozen sectioned rat kidney tissue labeling Ikaros with ab300405 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150081) at 1/1000 dilution (green). The nuclear counter stain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150081) at 1/1000 dilution.

No antigen retrieval was carried out.

Negative control (PMID: 1439790).

No staining on rat kidney.
Flow Cytometry (Intracellular) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Flow Cytometry (Intracellular) analysis of HeLa (human cervix adenocarcinoma epithelial cells, Left) / Jurkat (human T cell leukemia T lymphocytes, Right) labeling Ikaros (red) with ab300405 at a 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, (ab150081)) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730) Isotype control. Blue - (unlabeled control) - Cells without incubation with primary and secondary antibodies.

Negative control: HeLa (HPA data).
Flow Cytometry (Intracellular) - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Flow Cytometry (Intracellular) analysis of EL4 (mouse lymphoma T lymphocytes) labeling Ikaros (red) with ab300405 at a 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, (ab150081)) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730) Isotype control. Blue - (unlabeled control) - Cells without incubation with primary and secondary antibodies.
Immunoprecipitation - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Ikaros was immunoprecipitated from 10 μg Raji (human burkitt's lymphoma b lymphocytes) whole cell lysate with ab300405 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab300405 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1: Raji whole cell lysate 10 μg (Input).

Lane 2: Raji whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300405 in Raji whole cell lysate.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 8 seconds.

Observed MW(KDa): 30-70
Immunoprecipitation - Anti-Ikaros antibody [EPR25259-176] (AB300405)

Ikaros was immunoprecipitated from 10 μg EL4 (mouse lymphoma t lymphocytes) whole cell lysate with ab300405 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab300405 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1: EL4 whole cell lysate 10 μg (Input).

Lane 2: EL4 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300405 in EL4 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 8 seconds.

Observed MW(KDa): 30-70
Anti-Ikaros antibody [EPR25259-176] (AB300405)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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製品の概要

製品名

製品の詳細

由来種

特異性

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

組織特異性

関連疾患

配列類似性

ドメイン

翻訳後修飾

細胞内局在

参照データベース

製品の状態

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

Certificate of Compliance

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