NFKB1 antibody (AA 860-969)

Catalog No. ABIN561981

The Mouse Monoclonal anti-NFKB1 antibody (Clone 4A11) (ABIN561981) specifically detects NFKB1 in WB, ELISA, IF, IHC (p) and PLA. The antibody is reactive with Human samples. Independently validated for use in Immunofluorescence.

Quick Overview for NFKB1 antibody (AA 860-969) (ABIN561981)

Target: NFKB1 (Nuclear Factor of kappa Light Polypeptide Gene Enhancer in B-Cells 1 (NFKB1))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This NFKB1 antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Proximity Ligation Assay (PLA)

Clone: 4A11

Product Details anti-NFKB1 Antibody

Binding Specificity: AA 860-969

Purpose: Mouse monoclonal antibody raised against a partial recombinant NFKB1.

Sequence: MDNYEVSGGT VRELVEALRQ MGYTEAIEVI QAASSPVKTT SQAHSLPLSP ASTRQQIDEL RDSDSVCDSG VETSFRKLSF TESLTSGASL LTLNKMPHDY GQEGPLEGKI

Cross-Reactivity: Human

Characteristics: Antibody Reactive Against Recombinant Protein.

Immunogen: NFKB1 (AAH51765, 860 a.a. ~ 969 a.a) partial recombinant protein with GST tag. MW of the GST tag alone is 26 KDa.

Isotype: IgG1

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Independent Validation (IF)

Validation #104071 (Immunofluorescence)

by: Klinik für Anästhesiologie, Intensivmedizin und Schmerztherapie des Universitätsklinikums Knappschaftskrankenhaus Bochum

No.: #104071

Date: 11/21/2019

Antigen: NFKB1

Lot Number: 11067-4A11

Method validated: Immunofluorescence

Positive Control:

U-937 human monocytic cell line

Negative Control:

No primary antibody control

Notes:

Passed. ABIN561981 specifically stains NF-κB1 in U-937 human monocytic cells.

Validation Images

U937 monocytic cells stained with with anti-NF-κB1/p50 antibody ABIN561981 and an AF488-labeled secondary antibody (bottom row, green). Counterstain with DAPI (blue). Negative control without primary antibody (top row) shows no AF488 signal.

Full Methods

Primary Antibody: ABIN561981

Secondary Antibody: AF488-conjugated AffiniPure alpaca anti-mouse IgG antibody (Jackson Immuno Research, 615-545-214)

Full Protocol:

• Grow U-937 cells in RPMI 1640 Medium (Gibco, 21875034) supplemented with 10% FCS (Gibco, A3840001) and Pen/Strep (Gibco, 15140122), at 37°C and 5% CO₂ in 300µL in 24-well plates (Sarstedt AG&Co. KG).
• Use cytospin to spin cells onto slide.
• Fix cells on coverslide in 4% paraformaldehyde (Carl Roth GmbH, 0335.1) for 30min at RT.
• Wash cells 3x for 5min with PBS (Sigma-Aldrich, 8537).
• Permeabilize cells in 100µL PBS containing 0.1 % Triton X-100 (Sigma-Aldrich, T8787) for 5min at RT.
• Wash cells 3x for 5min with PBS.
• Block non-specific binding with 1x Duolink Blocking solution (Sigma-Aldrich, DUO82007) for 30min at RT.
• Incubate cells with primary mouse anti-human NF-κB1/p50 antibody (antibodies-online, ABIN561981, lot 11067-4A11) diluted 1:50 in 1x Duolink Antibody diluent (Sigma-Aldrich, DUO82008) ON at 4°C.
• Wash cells 3x for 5min with PBS.
• Incubate cells with secondary AF488-conjugated AffiniPure alpaca anti-mouse IgG antibody (Jackson Immuno Research, 615-545-214) diluted 1:400 in 1x Duolink Antibody diluent for 1h at RT in the dark.
• Wash cells 2x for 5min with PBS.
• Mount coverslips on glass slides in 50µL ProLong Gold antifade mountant with DAPI (Invitrogen, P36931).
• Image acquisition with an Olympus widefield microscope.

Experimental Notes:

Strong green staining dependent on the presence of primary antibody indicates that the antibody works. Negative control lacking the primary antibody shows only background fluorescence.

Handling

Buffer: In 1x PBS, pH 7.4

Handling Advice: Aliquot to avoid repeated freezing and thawing.

Storage: -20 °C

Storage Comment: Store at -20°C or lower. Aliquot to avoid repeated freezing and thawing.

Target Details for NFKB1

Target: NFKB1 (Nuclear Factor of kappa Light Polypeptide Gene Enhancer in B-Cells 1 (NFKB1))

Alternative Name: NFKB1

Background: Full Gene Name: nuclear factor of kappa light polypeptide gene enhancer in B-cells 1
Synonyms: DKFZp686C01211,EBP-1,KBF1,MGC54151,NF-kappa-B,NFKB-p105,NFKB-p50,p105,p50

Gene ID: 4790

Pathways: p53 Signaling, NF-kappaB Signaling, RTK Signaling, TCR Signaling, TLR Signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Myometrial Relaxation and Contraction, Regulation of Carbohydrate Metabolic Process, Hepatitis C, Toll-Like Receptors Cascades, BCR Signaling, S100 Proteins