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PRODUCT CODE: ET1609-78

Recombinant Phospho IKB alpha (S32) Monoclonal Antibody (ET1609-78)

  • Recombinant

Applications

  • WB

  • ICC

  • IP

REACTIVITY

  • Human

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Western blot analysis of Phospho-IKB alpha(S32) on SK-Br-3 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1609-78, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Phospho-IKB alpha(S32) on SK-Br-3 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1609-78, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of Phospho-IKB alpha(S32) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-78, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Western blot analysis of Phospho-IKB alpha(S32) on SK-Br-3 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1609-78, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IP

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Phospho IKB alpha (S32) Monoclonal Antibody (ET1609-78)

Immunogen

Recombinant protein

Host

Rabbit

Modification

Phospho

Modification Site

S32

Positive Control

SK-Br-3 cell lysates, Hela.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

ST53-05

PROPERTIES

Form

Liquid

Storage Condition

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

Storage Buffer

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

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

36 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC

  • 1:50-1:200

TARGET

UNIPROT #

P25963

PROTEIN NAME

Phospho-IKB alpha(S32)

SYNONYMS

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; NF kappa B inhibitor alpha antibody; NF-kappa-B inhibitor alpha antibody; NFKBI antibody; NFKBIA antibody; Nuclear factor of kappa light chain gene enhancer in B cells antibody; Nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor alpha antibody

SEQUENCE SIMILARITIES

Belongs to the NF-kappa-B inhibitor family.

POST-TRANSLATIONAL MODIFICATION

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.

SUBCELLULAR LOCATION

Cytoplasm, Nucleus.

FUNCTION

On the basis of both functional and structural considerations, members of the IκB family of proteins can be divided into four groups. The first of these groups, IκB-α, includes the avian protein pp40 and the mammalian MAD-3, both of which inhibit binding of p50-p65 NFkB complex or Rel protein to their cognate binding sites but do not inhibit the binding of p50 homodimer to κB sites, suggesting that the IκB-α family binds to the p65 subunit of p50-p65 heterocomplex through ankyrin repeats. The second member of the IκB family is represented by a protein designated IκB-β. The third group of IκB proteins is represented by IκB-γ, which is identical in sequence with the C-terminal domain of the p110 precursor of NFkB p50 and is expressed predominantly in lymphoid cells. An additional IκB family member, IκB-ε, has several phosphorylated forms and is primarily found complexed with Rel A and/or c-Rel.