Anti-phospho-NFKB p65 (Ser536) antibody
| 英文名称 | phospho-NFKB p65 (Ser536) |
| 中文名称 | 磷酸化细胞核因子抗体 |
| 别 名 | NF-kB p65 (phospho S536); p-NF-κB p65(Phospho-Ser536); RELA(phospho S536); NF kB P65; NF-kB p65; NFKBp65; NF-κBp65; p65 NF kappaB; p65 NFkB; NFKBp65; RELA; Transcription Factor p65; v rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B cells 3 (p65)); V Rel Avian Reticuloendotheliosis Viral Oncogene Homolog A; v rel reticuloendotheliosis viral oncogene homolog A (avian); v-rel reticuloendotheliosis viral oncogene homolog A; p65NFKB; Avian reticuloendotheliosis viral (v rel) oncogene homolog A; MGC131774; NFKB 3; NFKB3; Nuclear Factor NF Kappa B p65 Subunit; Nuclear factor of kappa light polypeptide gene enhancer in B cells 3; Nuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B Cells; TF65_HUMAN. |
DATASHEET
Host:Rabbit
Target Protein:phospho-NFKB p65 (Ser536)
IR:Immunogen Range:FS(p-S)IA
Clonality:Polyclonal
Isotype:IgG
Entrez Gene:5970
Swiss Prot:Q04206
Source:KLH conjugated Synthesised phosphopeptide derived from human NFKBp65 around the phosphorylation site of Ser536:FS(p-S)IA
Purification:affinity purified by Protein A
Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
Background: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].
Size:100ul
Concentration:1mg/ml
Applications:WB(1:500-2000)
IHC-P(1:100-500)
Flow-Cyt(1μg /test)
Cross Reactive Species:Human
Mouse
Rat
Chicken
Dog
Cow
Horse
.
For research use only. Not intended for diagnostic or therapeutic use.
VALIDATION IMAGES
Sample:
Lane 1: MCF-7 (Human) Cell Lysate at 30 ug
Lane 2: A431 (Human) Cell Lysate at 30 ug
Lane 3: HL60 (Human) Cell Lysate at 30 ug
Lane 4: Raw264.7 (Mouse) Cell Lysate at 30 ug
Lane 5: NIH/3T3 (Mouse) Cell Lysate at 30 ug
Lane 6: Jurkat (Human) Cell Lysate at 30 ug
Lane 7: A549 (Human) Cell Lysate at 30 ug
Primary: Anti-phospho-NFKB p65 (Ser536) (bs-0982R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 65 kD
Observed band size: 65 kD
Sample:
Lane 1: Large intestine (Mouse) Lysate at 40 ug
Lane 2: Large intestine (Rat) Lysate at 40 ug
Lane 3: Lung (Mouse) Lysate at 40 ug
Lane 4: Lung (Rat) Lysate at 40 ug
Lane 5: Bladder (Mouse) Lysate at 40 ug
Lane 6: RAW264.7 (Mouse) Cell Lysate at 30 ug
Primary: Anti-phospho-NFKB p65 (Ser536) (bs-0982R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 65 kD
Observed band size: 62 kD
Sample:
Lane 1: SiHa (Human) Cell Lysate at 30 ug
Lane 2: MCF-7 (Human) Cell Lysate at 30 ug
Lane 3: A431 (Human) Cell Lysate at 30 ug
Primary: Anti-phospho-NFKB p65 (Ser536) (bs-0982R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 65 kD
Observed band size: 62 kD
Sample:
Molt-4(Human) Cell Lysate at 30 ug
A549(Human) Cell Lysate at 30 ug
Jurkat(Human) Cell Lysate at 30 ug
Primary: Anti-phospho-NFKB p65 (Ser536) (bs-0982R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 65 kD
Observed band size: 62 kD
Sample:
Lung (Mouse) Lysate at 40 ug
Primary: Anti-phospho-NFKB p65(Ser536) (bs-0982R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 61 kD
Observed band size: 61 kD
Paraformaldehyde-fixed, paraffin embedded (mouse bladder); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-NFKB p65 (Ser536)) Polyclonal Antibody, Unconjugated (bs-0982R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human pancreatic cancer); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-NFKB p65 (Ser536)) Polyclonal Antibody, Unconjugated (bs-0982R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-NFKB p65 (Ser536)) Polyclonal Antibody, Unconjugated (bs-0982R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human breast); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-NFKB p65) Polyclonal Antibody, Unconjugated (bs-0982R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-NFKB p65 (Ser536)) Polyclonal Antibody, Unconjugated (bs-0982R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Blank control (blue line): HL60 (blue).
Primary Antibody (green line): Rabbit Anti-phospho-NFKB p65(Ser536) antibody (bs-0982R),Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1μg /test.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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