Human/Mouse IkB‑ alpha Antibody

Name: Human/Mouse IkB‑ alpha Antibody
Brand: R&D Systems
SKU: AF4299
Price: 189 USD
Availability: InStock

R&D Systems | Catalog # AF4299

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

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human, Mouse

Applications

Knockout Validated, Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

View all IkB-alpha Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse IkB-alpha
Met1-Leu314
Accession # Q9Z1E3

Specificity

Detects human and mouse IkB-alpha in Western blots.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human/Mouse IkB‑ alpha Antibody

Detection of Human/Mouse IkB-alpha by Western Blot.

Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line, DA3 mouse myeloma cell line, LNCaP human prostate cancer cell line, and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.1 µg/mL of Sheep Anti-Human/Mouse IkB-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for IkB-a at approximately 42 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

IkB‑ alpha in HeLa Human Cell Line.

IkB-a was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human/Mouse IkB-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

IkB‑ alpha in HeLa Human Cell Line.

IkB-a was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human/Mouse IkB-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western Blot Shows Human IkB‑ alpha Specificity by Using Knockout Cell Line.

Western blot shows lysates of HEK293T human embryonic kidney parental cell line and IkB-a knockout HEK293T cell line (KO). PVDF membrane was probed with 0.1 µg/mL of Sheep Anti-Human/Mouse IkB-a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for IkB-a at approximately 38 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of IkB‑ alpha in RAW 264.7 Mouse Monocyte/Macrophage Cell Line.

IkB‑ alpha was detected in immersion fixed RAW 264.7 Mouse Monocyte/Macrophage Cell Line using Sheep Anti-Human/Mouse IkB‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4299) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

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Applications for Human/Mouse IkB‑ alpha Antibody

Application

Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line and RAW 264.7 Mouse Monocyte/Macrophage Cell Line

Knockout Validated

IkB‑ alpha is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in IkB‑ alpha knockout HEK293T cell line.

Western Blot

0.1 µg/mL
Sample: NIH-3T3 mouse embryonic fibroblast cell line, DA3 mouse myeloma cell line, LNCaP human prostate cancer cell line, and Daudi human Burkitt's lymphoma cell line

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: IkB-alpha

NF kappa B inhibitor alpha (IkB-alpha ) inhibits the NF kappa B complex by binding and sequestering it in the cytoplasm. Upon stimulation, IkB-alpha is phosphorylated on serine residues marking it for degradation by the ubiquitin pathway. Following IkB-alpha degradation, the NF kappa B complex is able to translocate to the nucleus and activate transcription.