bsm-54207R [Primary Antibody]
NCK1 Recombinant Antibody
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Host: Rabbit

Target Protein: NCK1

Clonality: Recombinant

Isotype: IgG

Entrez Gene: 4690

Swiss Prot: P16333

Source: Recombinant protein corresponding human Nck1 aa 1-200

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol

Storage: Store at 4°C for up to 2 weeks. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.

Background:

Adapter protein which associates with tyrosine-phosphorylated growth factor receptors, such as KDR and PDGFRB, or their cellular substrates. Maintains low levels of EIF2S1 phosphorylation by promoting its dephosphorylation by PP1. Plays a role in the DNA damage response, not in the detection of the damage by ATM/ATR, but for efficient activation of downstream effectors, such as that of CHEK2. Plays a role in ELK1-dependent transcriptional activation in response to activated Ras signaling. Modulates the activation of EIF2AK2/PKR by dsRNA. May play a role in cell adhesion and migration through interaction with ephrin receptors.

Size: 100µL

Concentration: Lot Dependent

Applications: WB(1:300-5000)
FCM(1:20-100)
IHC-P(1:200-400)
IF(IHC-P)(1:50-200)
IF(ICC)(1:50-200)

Predicted Molecular Weight: 43/35


Predicted Cross Reactive Species: Human
Mouse
Rat

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

IF(ICC) staining with NCK1 (7C2) Monoclonal Antibody (bsm-54207R) at 1:100 in LOVO cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


IF(ICC) staining with NCK1 (7C2) Monoclonal Antibody (bsm-54207R) at 1:100 in A431 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


IF(ICC) staining with NCK1 (7C2) Monoclonal Antibody (bsm-54207R) at 1:100 in HUVEC cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with NCK1 Monoclonal Antibody, Unconjugated (bsm-54207R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with NCK1 Monoclonal Antibody, Unconjugated (bsm-54207R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with NCK1 Monoclonal Antibody, Unconjugated (bsm-54207R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Flow cytometric analysis of Jurkat cells with NCK1 (7C2) Monoclonal Antibody (bsm-54207R) 1:50 dilution (red) compared with an unlabeled control (cells without incubation with primary antibody; black).


The Hela (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-NCK1 antibody (bsm-54207R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.


Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with NCK1 Monoclonal Antibody, Unconjugated (bsm-54207R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with NCK1 Monoclonal Antibody, Unconjugated (bsm-54207R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with NCK1 Monoclonal Antibody, Unconjugated (bsm-54207R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.


25 ug total protein per lane of various lysates (see on figure) probed with NCK1 monoclonal antibody, unconjugated (bsm-54207R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.