IkB-alpha Antibody (6A920) - Azide and BSA Free

Name: IkB-alpha Antibody (6A920) - Azide and BSA Free
Brand: Novus Biologicals
SKU: NBP2-80787
Price: 419 USD
Availability: InStock

Novus Biologicals | Catalog # NBP2-80787

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Product Specifications
Scientific Data
Applications
Flow Cytometry
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Human, Mouse, Rat

Applications

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ kappa Clone # 6A920

Format

Azide and BSA Free

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Product Specifications

Immunogen

A recombinant protein corresponding to amino acid residues 32-291 of human IkBa was used as immunogen.

Reactivity Notes

Rat reactivity reported in the scientific literature (PMID: 23840265).

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁ kappa

Scientific Data Images for IkB-alpha Antibody (6A920) - Azide and BSA Free

Western Blot: IkB-alpha Antibody (6A920)Azide and BSA Free [NBP2-80787]

Western Blot: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - Lysates of Jurkat human acute T cell leukemia cell line, LNCaP human prostate cancer cell line, PCx2011;3 human prostate cancer cell line, HeLa human cervical epithelial carcinoma cell line, and NIHx2011;3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 0.5 ug/mL mouse anti-IkB-a monoclonal (NB100-56507, Novus Biologicals), followed by 1:2000 dilution of the appropriate HRP-conjugated secondary antibody, donkey anti-mouse IgG. Image from the standard format of this antibody.

Immunocytochemistry/ Immunofluorescence: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787]

Immunocytochemistry/Immunofluorescence: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - Hek293 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-IkB-alpha (6A920) at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective. Image from the standard format of this antibody.

Immunohistochemistry: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787]

Immunohistochemistry: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - Human breast probed with IkBa antibody at 5 ug/ml. Human tissue TMA was used for this test. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min

Flow Cytometry: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787]

Flow Cytometry: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - Intracellular staining of 10^6 ThP-1 cells using 0.25 ug of NB100-56507. Shaded histogram represents cells alone, green represents the isotype control, and red represents the IkBa antibody. Novus's intracellular flow kit was used for this test, and an anti-mouse IgG FITC conjugated secondary. Image from the standard format of this antibody.

Western Blot: IkB-alpha Antibody (6A920)Azide and BSA Free [NBP2-80787]

Western Blot: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - IkBa using NB100-56507 at 2 ug/ml in (A) Daudi and (B) NIH 3T3 whole cell lysate. Image from the standard format of this antibody.

Western Blot: IkB-alpha Antibody (6A920)Azide and BSA Free [NBP2-80787]

Western Blot: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - Jurkat cells were treated for 30 min with 100 ug/ml ALLN (N-Acetyl-Leu-Leu-Norleucinal; a Calpain inhibitor and also proteasome inhibitor that prevents IkBa dephosphorylation) followed by incubation with (lanes 2 & 4) or without 1 nM TNF-a (1 & 3). The membranes were blotted with NB100-56724 (lanes 1 & 2) or NB100-56507 (that recognizes both non-phospho and phosphorylated forms of IkBa) and immunoreactivity was detected by ECL. The data shows that NB100-56724 detects specifically the phosphorylated form of IkBa. Image from the standard format of this antibody.

Flow Cytometry: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787]

Flow Cytometry: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - An intracellular stain was performed on Hek293 cells with IkB-alpha Antibody [6A920] NB100-56507 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550. Image from the standard format of this antibody.

Flow Cytometry: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787]

Flow Cytometry: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - An intracellular stain was performed on NIH3T3 cells with IkB-alpha Antibody [6A920] NB100-56507 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550. Image from the standard format of this antibody.

Simple Western: IkB-alpha Antibody (6A920)Azide and BSA Free [NBP2-80787]

Simple Western: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - Simple Western lane view shows a specific band for IKB alpha in 0.5 mg/ml of NIH-3T3 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Image from the standard format of this antibody.

Western Blot: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787]

Western Blot: IkB-alpha Antibody (6A920) - Azide and BSA Free [NBP2-80787] - Western blot shows lysates of HEK293 human embryonic kidney parental cell line and IkB-alpha knockout (KO) HEK293 human embryonic kidney cell line. PVDF membrane was probed with 2 ug/ml of Mouse Anti-Human IkB-alpha monoclonal Antibody (Catalog # NB100-56507) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody. Specific band was detected for IkB-alpha at approximately 38 kDa (as indicated) in the parental HEK293 celll line, but is not detectable in the knockout HEK293 cell line. This experiment was conducted under reducing conditions. Image from the standard format of this antibody.

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Applications for IkB-alpha Antibody (6A920) - Azide and BSA Free

Application

Recommended Usage

Flow Cytometry

0.25-1 ug/10^6 cells

Immunocytochemistry/ Immunofluorescence

1:20-1:1000

Immunohistochemistry

1:20-1:1000

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

1 ug/ml

Simple Western

1:20

Western Blot

1-2 ug/ml

Application Notes

In Simple Western only 10 - 15 ul of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: antibody dilution of 1:20. This antibody is CyTOF ready.

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Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: IkB-alpha

IkBa Inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses IkBa becomes phosphorylated, promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription. IkB proteins are phosphorylated by IkB kinase complex consisting of at least three proteins, IKK1/a, IKK2/b, and IKK3/g.

Long Name

I-kappa-B-alpha

Alternate Names

IkBalpha, MAD-3, NFKBIA

Gene Symbol

NFKBIA

Additional IkB-alpha Products

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Product Specific Notices for IkB-alpha Antibody (6A920) - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.