WB
Cell treatment (CT) Western blot analysis of Phospho-IKB alpha (S32/S36) on different lysates with Rabbit anti-Phospho-IKB alpha (S32/S36) antibody at 1/5,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 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 1/5,000 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.IHC
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Phospho-IKB alpha (S32/S36) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 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.ICC/IF
Cell treatment (CT) Immunocytochemistry analysis of HeLa cells treated with 20ng/mL TNF-α and 100nM Calyculin A for 10 minutes labeling Phospho-IKB alpha (S32/S36) with Rabbit anti-Phospho-IKB alpha (S32/S36) antibody at 1/1,000 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/S36) antibody at 1/1,000 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.ICC/IF
Cell treatment (CT) Immunocytochemistry analysis of NIH/3T3 cells treated with 20ng/mL TNF-α and 100nM Calyculin A for 10 minutes labeling Phospho-IKB alpha (S32/S36) with Rabbit anti-Phospho-IKB alpha (S32/S36) 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/S36) 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/S36) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser32/Ser36 of human IKB alpha aa 1-50. |
| Modification | p-S32/S36 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHCWB |
| WB:1:5000 IHC:1:200 ICC/IF:1:100-1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | NFKBIA |
|---|---|
| Gene Synonyms | IKBA MAD-3 NFKBI EDAID2 |
| Gene Full Name | NFKB inhibitor alpha |
| Gene Summary | This gene encodes a member of the NF-kappa-B inhibitor family, which contain multiple ankrin repeat domains. The encoded protein interacts with REL dimers to inhibit NF-kappa-B/REL complexes which are involved in inflammatory responses. The encoded protein moves between the cytoplasm and the nucleus via a nuclear localization signal and CRM1-mediated nuclear export. Mutations in this gene have been found in ectodermal dysplasia anhidrotic with T-cell immunodeficiency autosomal dominant disease. [provided by RefSeq, Aug 2011] |
| 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 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) | 36 kDa |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Cell treatment (CT) Western blot analysis of Phospho-IKB alpha (S32/S36) on different lysates with Rabbit anti-Phospho-IKB alpha (S32/S36) antibody at 1/5,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 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 1/5,000 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.
IHC
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Phospho-IKB alpha (S32/S36) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 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.
ICC/IF
Cell treatment (CT) Immunocytochemistry analysis of HeLa cells treated with 20ng/mL TNF-α and 100nM Calyculin A for 10 minutes labeling Phospho-IKB alpha (S32/S36) with Rabbit anti-Phospho-IKB alpha (S32/S36) antibody at 1/1,000 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/S36) antibody at 1/1,000 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.
ICC/IF
Cell treatment (CT) Immunocytochemistry analysis of NIH/3T3 cells treated with 20ng/mL TNF-α and 100nM Calyculin A for 10 minutes labeling Phospho-IKB alpha (S32/S36) with Rabbit anti-Phospho-IKB alpha (S32/S36) 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/S36) 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.| Application Notes | WB:1:5000 IHC:1:200 ICC/IF:1:100-1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
