MA5-16157

antibody from Invitrogen Antibodies

Targeting: CHUK IkBKA, IKK-alpha, IKK1, IKKA, NFKBIKA, TCF16

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Recommended by provider)

Antibody Data

Product number

MA5-16157

Provider

Invitrogen Antibodies

Product name

IKK alpha Monoclonal Antibody (14A231)

Antibody type

Monoclonal

Antigen

Other

Description

Suggested positive control: Daudi, NIH-3T3, or Raw.

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

14A231

Vial size

100 µg

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of IKK alpha in Daudi cell lysate using a IKK alpha monoclonal antibody (Product # MA5-16157) at 1 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of IKK alpha in 0.5 mg/mL Daudi (left) and U937 (right) lysate. Samples were incubated in IKK alpha monoclonal antibody (Product # MA5-16157). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of IKK alpha in total protein from various human, mouse and Rat cell lines. Samples were incubated in IKK alpha monoclonal antibody (Product # MA5-16157) using a dilution of 1.0 µg/mL followed by an anti-mouse HRP secondary antibody. Separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. Detection: chemiluminescence.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of IKK alpha in Daudi cell lysate. Samples were incubated in IKK alpha monoclonal antibody (Product # MA5-16157) using a dilution of 1 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of IKK alpha was performed by loading 30 µg of ME180 (lane1), ME180- IKK alpha knockout (lane2) whole cell lysate using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. IKK alpha was detected at ~ 80 kDa using IKK alpha Mouse Monoclonal Antibody (Product # MA5-16157) at 1 µg/mL in 5% skim milk at 4°C overnight on a rocking platform. Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody HRP conjugate (Product # A28177, 1:4000 dilution) was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to IKK alpha

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of IKK alpha in 0.5 mg/mL Daudi (left) and U937 (right) lysate. Samples were incubated in IKK alpha monoclonal antibody (Product # MA5-16157). This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of IKK alpha in formalin-fixed paraffin-embedded tissue section of human kidney. Samples were incubated in IKK alpha monoclonal antibody (Product # MA5-16157) using a dilution of 1:200 followed by HRP labeled anti-rabbit secondary antibody and DAB reagent. Nuclei of cells were counter-stained with hematoxylin.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Intracellular staining of HEK293 cells using 0.5 µg of IKK alpha monoclonal antibody (Product # MA5-16157). Green histogram represents the isotype control antibody, red represents the IKK alpha monoclonal antibody (Product # MA5-16157). anti-mouse IgG1 PE conjugated secondary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of IKK alpha in HEK293 cells. Samples were incubated in IKK alpha monoclonal antibody (Product # MA5-16157) using a dilution of 2.5 µg/mL for 30 minutes at room temperature followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (Product # 35503). Antibody (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin.