IKK alpha + IKK beta Recombinant Rabbit Monoclonal Antibody [SN63-02]

Overview

Product Name

IKK alpha + IKK beta Recombinant Rabbit Monoclonal Antibody [SN63-02] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human IKK beta aa 450-652 / 756.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IP

Target Molecular Weight

Predicted band size: 87 kDa

Positive Control

HeLa cell lysate, 293T cell lysate, Daudi cell lysate, A431, NIH/3T3, C6.

Conjugation

unconjugated

Clone Number

SN63-02

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

WB
1:2,000

IF-Cell
1:100-1:500

IP
Use at an assay dependent concentration.

Target

Function

IKK-α is part of the IκB kinase complex that plays an important role in regulating the NF-κB transcription factor. IKK-β is an enzyme that serves as a protein subunit of IκB kinase, which is a component of the cytokine-activated intracellular signaling pathway involved in triggering immune responses. IKK's activity causes activation of a transcription factor known as Nuclear Transcription factor kappa-B or NF-κB. Activated IKK-β phosphorylates a protein called the inhibitor of NF-κB, IκB (IκBα), which binds NF-κB to inhibit its function. IKK-α and IKK-β phosphorylate the IκB proteins, marking them for degradation via ubiquitination and allowing NF-κB transcription factors to go into the nucleus. Once activated, NF-κB transcription factors regulate genes that are implicated in many important cellular processes, including immune response, inflammation, cell death, and cell proliferation.

Background References

1. Hwang, B. et al. 2015. IPO3-mediated Nonclassical Nuclear Import of NF-κB Essential Modulator (NEMO) Drives DNA Damage-dependent NF-κB Activation. The Journal of biological chemistry. 290: 17967-84.

2. Muhammad, K. et al. 2014. NF-κB factors control the induction of NFATc1 in B lymphocytes. European journal of immunology. 44: 3392-402.

Subcellular Location

Cytoplasm, Nucleus, Membrane raft.

UNIPROT

SwissProt: O14920 Human

SwissProt: O15111 Human

SwissProt: O88351 Mouse

SwissProt: Q60680 Mouse

SwissProt: Q9QY78 Rat

Synonyms

CHUK antibody

Conserved helix loop helix ubiquitous kinase antibody

I kappa B kinase 1 antibody

I kappa B kinase 2 antibody

I Kappa B kinase alpha antibody

I Kappa B kinase beta antibody

IkB kinase alpha subunit antibody

IkBKA antibody

IkBKB antibody

IKK a kinase antibody

Expand

Images

Western blot analysis of IKK alpha + IKK beta on different lysates with Rabbit anti-IKK alpha + IKK beta antibody (HA750248) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: 293T cell lysate
Lane 3: Daudi cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 87 kDa
Observed band size: 87 kDa

Exposure time: 1 minute 2 seconds; ECL: K1802;

4-20% 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 (HA750248) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)

Western blot analysis of IKK alpha + IKK beta on different lysates with Rabbit anti-IKK alpha + IKK beta antibody (HA750248) at 1/5,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-IKK beta KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 87 kDa
Observed band size: 87 kDa

Exposure time: 30 seconds; ECL: K1801;

4-20% 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 (HA750248) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of A431 cells labeling IKK alpha + IKK beta with Rabbit anti-IKK alpha + IKK beta antibody (HA750248) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-IKK alpha + IKK beta antibody (HA750248) at 1/500 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 (HA601187, 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 IKK alpha + IKK beta with Rabbit anti-IKK alpha + IKK beta antibody (HA750248) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-IKK alpha + IKK beta antibody (HA750248) at 1/500 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 (HA601187, 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 C6 cells labeling IKK alpha + IKK beta with Rabbit anti-IKK alpha + IKK beta antibody (HA750248) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-IKK alpha + IKK beta antibody (HA750248) at 1/250 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 (HA601187, 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"