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Immunofluorescence analysis of I-kappa-B-alpha was done on 70% confluent log phase NIH/3T3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with I-kappa-B-alpha(ZI002) Mouse Monoclonal Antibody (397700) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Cytoplasmic and Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Mouse, Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Synthetic peptide derived from the C-terminal region of the human I-kappa-B-alpha protein|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
NFKB1 or NFKB2 is bound to REL, RELA, or RELB to form the NFKB complex. The NFKB complex is inhibited by I-kappa-B proteins (NFKBIA or NFKBIB, MIM 604495), which inactivate NF-kappa-B by trapping it in the cytoplasm. Phosphorylation of serine residues on the I-kappa-B proteins by kinases (IKBKA, MIM 600664, or IKBKB, MIM 603258) marks them for destruction via the ubiquitination pathway, thereby allowing activation of the NF-kappa-B complex. Activated NFKB complex translocates into the nucleus and binds DNA at kappa-B-binding motifs such as 5-prime GGGRNNYYCC 3-prime or 5-prime HGGARNYYCC 3-prime (where H is A, C, or T; R is an A or G purine; and Y is a C or T pyrimidine).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
||KU812 cells provide a novel in vitro model of the human IL-33/ST2L axis: functional responses and identification of signaling pathways.||Tare N,Li H,Morschauser A,Cote-Sierra J,Ju G,Renzetti L,Lin TA||Experimental cell research (316:2527)||2010|
|Human||1 ug/ml||KU812 cells provide a novel in vitro model of the human IL-33/ST2L axis: functional responses and identification of signaling pathways.||Tare N,Li H,Morschauser A,Cote-Sierra J,Ju G,Renzetti L,Lin TA||Experimental cell research (316:2527)||2010|
I(Kappa)B(alpha); I-kappa-B-alpha; I-kappaBalpha; IkappaBalpha; ikB-alpha; IKBA; MAD3; major histocompatibility complex enhancer-binding protein MAD3; NF-kappa-B inhibitor alpha; NF-kappaB inhibitor alpha; NFKBI; NFKBIA; nuclear factor of kappa light chain gene enhancer in B-cells; nuclear factor of kappa light chain gene enhancer in B-cells inhibitor, alpha; nuclear factor of kappa light polyp gene enhancer in B-cell 1; nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha
AI462015; IKBA; MAD-3; MAD3; NFKBI; NFKBIA