WB
Western blot analysis of Phospho-IKB alpha (S32) on different lysates with Rabbit anti-Phospho-IKB alpha (S32) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HeLa treated with 20ng/mL TNF-α for 5 minutes cell lysate, Lane 3: HeLa treated with 20ng/mL TNF-α for 5 minutes cell lysate, then the membrane treated with λpp for 1 hour, Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute 2 seconds; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Phospho-IKB alpha (S32) with Rabbit anti-Phospho-IKB alpha (S32) antibody 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-Phospho-IKB alpha (S32) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.| Product Name | Phospho-IKB alpha (S32) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | 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. |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser32 of human IKB alpha. |
| Modification | p-S32 |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFWB |
| WB:1:1,000 ICC/IF:1:50-1:200 |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | NFKBIA |
|---|---|
| Gene Synonyms | IKBA MAD-3 NFKBI EDAID2 |
| Gene Full Name | NFKB inhibitor alpha |
| Alternative Names | 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 |
| Molecular Weight(MW) | 36kDa |
| Cellular Localization | Cytoplasm, Nucleus. |
| SwissProt ID | P25963 |
|---|
WB
Western blot analysis of Phospho-IKB alpha (S32) on different lysates with Rabbit anti-Phospho-IKB alpha (S32) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate, Lane 2: HeLa treated with 20ng/mL TNF-α for 5 minutes cell lysate, Lane 3: HeLa treated with 20ng/mL TNF-α for 5 minutes cell lysate, then the membrane treated with λpp for 1 hour, Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute 2 seconds; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Phospho-IKB alpha (S32) with Rabbit anti-Phospho-IKB alpha (S32) antibody 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-Phospho-IKB alpha (S32) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.| Positive Control | Ags, NIH/3T3, Hela. |
|---|---|
| Application Notes | WB:1:1,000 ICC/IF:1:50-1:200 |
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
