Human Total IkB-alpha DuoSet IC ELISA, 2 Plate

Lysate prepared from MCF-7 human breast cancer cells was incubated in wells coated with Total I kappa B-alpha Capture Antibody. Unbound material was removed by washing and bound material was solubilized in SDS gel sample buffer. The same lysate and captured proteins were resolved by SDS-PAGE, transferred to a PVDF membrane, and immunoblotted with Total I kappa B-alpha Detection Antibody. In addition to IgG heavy and light chains, only a single band at 39 kDa corresponding to I kappa B-alpha was detected in captured material, indicating that the ELISA is specific for I kappa B-alpha.Specificity was further demonstrated using cross-reactivity experiments with related I kappa B family members. Recombinant human (rh) I kappa B-beta and rhI kappa B-epsilon were tested at 50 ng/mL and did not cross-react or interfere in the assay.

Lysates prepared from MCF-7 human breast cancer, Daudi human Burkitt's lymphoma, and HeLa human cervical epithelial carcinoma cells were quantified with this DuoSet® IC ELISA. The same lysates were also immunoblotted (inset) with anti-I kappa B-alpha monoclonal antibody (Catalog # MAB4299). The DuoSet® IC ELISA results correlate well with the relative amounts of I kappa B-alpha detected by Western Blot.

Lysates prepared from HeLa human cervical epithelial carcinoma cells either untreated or treated with 20 ng/mL of recombinant human TNF- alpha (Catalog # 210-TA) for 20 minutes were quantified with this DuoSet® IC ELISA. The same lysates were also immunoblotted (inset) with anti-I kappa B-beta monoclonal antibody. The DuoSet® IC ELISA results correlate well with the relative amounts of I kappa B-alpha detected by Western Blot.

• Reactivity: Hu
• Applications: ELISA
• Conjugate: Biotin

Human Total IkB-alpha DuoSet IC ELISA, 2 Plate Summary

• Source: N/A
• Assay Type: Solid Phase Sandwich ELISA
• Inter-Assay: See PDF Datasheet for details
• Intra-Assay: See PDF Datasheet for details
• Spike Recovery: See PDF Datasheet for details
• Sample Volume: See PDF Datasheet for details
• Gene: NFKBIA

Applications/Dilutions

• Dilutions:
  • ELISA
• Application Notes: No significant interference observed with available related molecules.

Packaging, Storage & Formulations

• Storage: Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Human Total IkB-alpha DuoSet IC ELISA, 2 Plate

• alpha
• I-kappa-B-alpha
• IkappaBalphaNF-kappa-B inhibitor alpha
• IKBAikB-alpha
• IkBalpha
• IkB-alpha
• MAD3
• MAD-3
• Major histocompatibility complex enhancer-binding protein MAD3
• 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

Background

The inhibitory IkB proteins (alpha, beta, epsilon) complex with and sequester NFkB family members in the cytoplasm, rendering these transcription factors inactive. When phosphorylated by an active IkB kinase (IKK) complex, IkB proteins are degraded by proteasomes, resulting in the release and nuclear translocation of active NFkB.

Publications for IkB-alpha (DYC4299-2)(2)

Publications using DYC4299-2

• Piao, CH;Fan, Y;Nguyen, TV;Song, CH;Kim, HT;Chai, OH; PM2.5 exposure regulates Th1/Th2/Th17 cytokine production through NF-?B signaling in combined allergic rhinitis and asthma syndrome International immunopharmacology 2023-05-08 [PMID: 37163921] (Mouse)
• JH Liu, CH Hsieh, CY Liu, CW Chang, YJ Chen, TH Tsai Anti-inflammatory effects of Radix aucklandiae herbal preparation ameliorate intestinal mucositis induced by 5-fluorouracil in mice Journal of ethnopharmacology, 2021-02-07;271(0):113912. 2021-02-07 [PMID: 33567307] (Mouse)

FAQs for IkB-alpha (DYC4299-2). (Showing 1 - 1 of 1 FAQs).

1. What is the difference between the Quantikine and Duoset ELISA kits?
   • Our Quantikine kits are fully optimized and validated for the sample types listed on the product specific webpage and datasheet, as this can vary between kits. Each kit is supplied ready to use with one pre-coated 96-well plate, a detection antibody directly conjugated to HRP, and all other necessary reagents. These kits are ideal for researchers who want the convenience of a ready to use and optimized ELISA product. Some of these kits are also available prepackaged in larger 6 and 50 plate sizes.Our DuoSet Kits, in contrast, are ELISA development kits containing the capture and detection antibody, the mass-value calibrated standard, and streptavidin-HRP to prepare approximately 5 or 15 plates. Ancillary reagents will need to be used/purchased, and for most kits, we will recommend one of our Ancillary Reagent Kits, which contain the reagents we use ourselves in-house. DuoSet kits are validated only for cell culture supernatant samples and therefore require further development and validation for accurate measurement in more complex samples such as serum and plasma. Our DuoSet Kits offer an economical, flexible alternative for the experienced ELISA user.

Bioinformatics

• Gene Symbol: NFKBIA