IKK alpha + IKK beta Recombinant Rabbit Monoclonal Antibody [SN63-02] - BSA and Azide free
Catalog# HA750248
IKK alpha + IKK beta Recombinant Rabbit Monoclonal Antibody [SN63-02] - BSA and Azide free
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WB
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IF-Cell
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IP
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Human
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Mouse
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Rat
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ET1611-23
含抗保成分
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unconjugated
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
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
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WB
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1:2,000
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IF-Cell
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1:100-1:500
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IP
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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
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
ExpandImages
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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"
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