NF-KB p65 Antibody (YA268)

NF-KB p65 Antibody is a non-conjugated and Rabbit origined monoclonal antibody about 60 kDa, targeting to NF-KB p65. It can be used for WB,IHC-F,IHC-P,ICC/IF,IP assays with tag free, in the background of Human, Mouse.

NF-KB p65: NF-kappa-B is a ubiquitous transcription factor involved in several biological processes. It is held in the cytoplasm in an inactive state by specific inhibitors. Upon degradation of the inhibitor, NF-kappa-B moves to the nucleus and activates transcription of specific genes. NF-kappa-B is composed of NFKB1 or NFKB2 bound to either REL, RELA, or RELB. The most abundant form of NF-kappa-B is NFKB1 complexed with the product of this gene, RELA. Four transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Sep 2011]

Free

Q04206

Predicted band size: 60 kDa; Observed band size: 65 kDa

affinity purified

Non-conjugated

Unmodified

AB_3102184

Cell Biology

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:500-1:1000 ;IHC: 1:50-1:100 ;IF: 1:50-1;200 ;IP: 1:20

• 
  Western blot analysis of extracts from K562 (lane 2(20μg), RAW264.7 (lane 3(20μg) and Hela (lane 4(20μg) using NF-KB p65 (HY-P80765) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of Hela cells labeling NF-KB p65 with NF-KB p65 antibody (HY-P80765) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with NF-KB p65 antibody (HY-P80765) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of NIH/3T3 cells labeling NF-KB p65 with NF-KB p65 antibody (HY-P80765) at 1/50 dilution. Confocal image showing the signal translocated from the cytoplasm into the nucleus in NIH/3T3 cells after the treatment with TNF-alpha (50 ng/ml) for 20 min. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with NF-KB p65 antibody (HY-P80765) at 1/200 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

WB, IHC-F, IHC-P, ICC/IF, IP

Liquid

Supplied in 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol and 0.05% BSA. Preservative: 0.01% Sodium azide

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human NF-kB p65 aa500-551.