Phospho-IKB alpha (S32) Recombinant Rabbit Monoclonal Antibody [ST53-05]
cat.: ET1609-78
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell, IP |
| Clonality: | Monoclonal |
| Clone number: | ST53-05 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 36 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser32 of human IKB alpha. |
| Positive control: | HeLa treated with 20ng/mL TNF-α for 5 minutes cell lysate, NIH/3T3 cells were starved for 18 hours then treated with 20 ng/ml TNF alpha for 5 minutes, HeLa. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IF-Cell IP |
1:1,000 1:50-1:200 Use at an assay dependent concentration. |
| Uniprot #: | SwissProt: P25963 Human | Q9Z1E3 Mouse |
| Alternative names: | I kappa B alpha I-kappa-B-alpha IkappaBalpha IkB-alpha IKBA IKBA_HUMAN IKBalpha MAD 3 MAD3 Major histocompatibility complex enhancer-binding protein MAD3 NF kappa B inhibitor alpha NF-kappa-B inhibitor alpha NFKBI NFKBIA Nuclear factor of kappa light chain gene enhancer in B cells Nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor alpha |
Images
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Fig1:
Western blot analysis of Phospho-IKB alpha (S32) on different lysates with Rabbit anti-Phospho-IKB alpha (S32) antibody (ET1609-78) 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. Predicted band size: 36 kDa Observed band size: 36 kDa 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 (ET1609-78) at 1/1,000 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. |
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Fig2:
Western blot analysis of Phospho-IKB alpha (S32) on NIH-3T3 cell lysates. Lane 1: NIH/3T3 cells were starved for 18 hours, whole cell lysate, 10 ug/lane. Lane 2: NIH/3T3 cells were starved for 18 hours, then treated with 20 ng/ml TNF alpha for 5 minutes, whole cell lysates, 10 ug/lane. Lane 3: NIH/3T3 cells were starved for 18 hours, then treated with 20 ng/ml TNF alpha for 10 minutes, whole cell lysates, 10 ug/lane. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody Anti-Phospho-IKB alpha (S32) (ET1609-78, 1/1,000) , Anti-IKB alpha antibody ( ET1603-6, 1/2,000) and Anti-GAPDH antibody (ET1601-4, 1/10,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Predicted band size: 36 kDa Observed band size: 36 kDa Exposure time: 1 minute |
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Fig3:
Immunocytochemistry analysis of HeLa cells labeling Phospho-IKB alpha (S32) with Rabbit anti-Phospho-IKB alpha (S32) antibody (ET1609-78) 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 (ET1609-78) at 1/100 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 (M1305-2, 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.