IKB alpha Recombinant Rabbit Monoclonal Antibody [SZ00-07]

RMB: 900 特惠 1500 1500

产品规格

50μl

100μl

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Catalog# ET1603-6

IKB alpha Recombinant Rabbit Monoclonal Antibody [SZ00-07]

Application

WB
IF-Cell
IF-Tissue
IHC-P
IP
FC

Reactivity

Human
Mouse
Rat

This product has been cited in peer reviewed publications, see list HERE

概述

产品名称

IKB alpha Recombinant Rabbit Monoclonal Antibody [SZ00-07]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Synthetic peptide within human IKB alpha aa 1-50.

种属反应性

Human, Mouse, Rat

验证应用

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC

分子量

Predicted band size: 36 kDa

阳性对照

HeLa cell lysate, SK-Br-3 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, Hela, MCF-7, SHG-44, human kidney tissue, mouse kidney tissue, mouse brain tissue, mouse stomach tissue.

偶联

unconjugated

克隆号

SZ00-07

RRID

AB_3065034

产品特性

形态

Liquid

存放说明

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

存储缓冲液

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

WB
1:5,000-1:20,000

IF-Cell
1:100-1:200

IF-Tissue
1:200-1:500

IHC-P
1:200-1:1,000

IP
Use at an assay dependent concentration.

FC
1:1,000

发表文章中的应用

WB	查看 46 篇文献如下
IHC-P	查看 3 篇文献如下

发表文章中的种属

Mouse	查看 29 篇文献如下
Human	查看 15 篇文献如下
Rat	查看 4 篇文献如下
Zebrafish	查看 1 篇文献如下

靶点

功能

NFKB1 or NFKB2 is bound to REL, RELA, or RELB to form the NFKB complex. The NFKB complex is inhibited by I-kappa-B proteins, which inactivate NF-kappa-B by trapping it in the cytoplasm. Phosphorylation of serine residues on the I-kappa-B proteins by kinases marks them for destruction via the ubiquitination pathway, thereby allowing activation of the NF-kappa-B complex. Activated NFKB complex translocates into the nucleus and binds DNA at kappa-B-binding motifs such as 5-prime GGGRNNYYCC 3-prime or 5-prime HGGARNYYCC 3-prime.

背景文献

1. Bai X et al. Prostaglandin E2 stimulates 1-integrin expression in hepatocellular carcinoma through the EP1 receptor/PKC/NF- B pathway. Sci Rep 4:6538 (2014).

2. Broderick TL et al. Downregulation in GATA4 and Downstream Structural and Contractile Genes in the db/db Mouse Heart. ISRN Endocrinol 2012:736860 (2012).

序列相似性

Belongs to the NF-kappa-B inhibitor family.

翻译后修饰

Phosphorylated; disables inhibition of NF-kappa-B DNA-binding activity. Phosphorylation at positions 32 and 36 is prerequisite to recognition by UBE2D3 leading to polyubiquitination and subsequent degradation.; Sumoylated; sumoylation requires the presence of the nuclear import signal. Sumoylation blocks ubiquitination and proteasome-mediated degradation of the protein thereby increasing the protein stability.; Monoubiquitinated at Lys-21 and/or Lys-22 by UBE2D3. Ubiquitin chain elongation is then performed by CDC34 in cooperation with the SCF(FBXW11) E3 ligase complex, building ubiquitin chains from the UBE2D3-primed NFKBIA-linked ubiquitin. The resulting polyubiquitination leads to protein degradation. Also ubiquitinated by SCF(BTRC) following stimulus-dependent phosphorylation at Ser-32 and Ser-36.; Deubiquitinated by porcine reproductive and respiratory syndrome virus Nsp2 protein, which thereby interferes with NFKBIA degradation and impairs subsequent NF-kappa-B activation.

亚细胞定位

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P25963 Human

SwissProt: Q9Z1E3 Mouse

SwissProt: Q63746 Rat

别名

I kappa B alpha antibody

I-kappa-B-alpha antibody

IkappaBalpha antibody

IkB-alpha antibody

IKBA antibody

IKBA_HUMAN antibody

IKBalpha antibody

MAD 3 antibody

MAD3 antibody

Major histocompatibility complex enhancer-binding protein MAD3 antibody

展开

图片

Western blot analysis of IKB alpha on different lysates with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa cell lysate
Lane 3: SK-Br-3 cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: RAW264.7 cell lysate
Lane 6: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 1 minute;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1603-6) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
IKB alpha was immunoprecipitated from 0.5 mg Hela whole cell lysates with ET1603-6 at 2 μg/mL. Western blot was performed from the immunoprecipitate using ET1603-6 at 1/5,000 dilution for 45 minutes at room temperature. Goat anti-Rabbit IgG-HRP Secondary Antibody (HA1001) was used at 1:300,000 dilution for 30 minutes at room temperature.

Lane 1: Hela whole cell lysates at 10 μg;
Lane 2: IKB alpha (ET1603-6) IP in Hela whole cell lysates;
Lane 3: Rabbit IgG instead of IKB alpha (ET1603-6) in Hela whole cell lysates.

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 1 minute;

12% SDS-PAGE gel.
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1603-6) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1603-6) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1603-6) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunocytochemistry analysis of HeLa cells labeling IKB alpha with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of NIH/3T3 cells labeling IKB alpha with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of PC-12 cells labeling IKB alpha with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IKB alpha antibody (ET1603-6) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Flow cytometric analysis of HeLa cells labeling IKB alpha.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1603-6, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).