Anti-CCR7 Polyclonal Antibody is a Rabbit antibody targeting CCR7. Anti-CCR7 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog)

1. Blank control (Black line): Mouse spleen (Black).
Primary Antibody (green line): Rabbit Anti-CD4 antibody (TMAB-00326-PE)
Dilution: 3 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG-PE.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
2. Tissue/cell: human laryngocarcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-CCR7 Polyclonal Antibody, Unconjugated (TMAB-00326) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Tissue/cell: human gastric tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-CCR7 Polyclonal Antibody, Unconjugated (TMAB-00326) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, FITC conjugated used at 1:200 dilution for 40 minutes at 37°C. DAPI (5 μg/ml,blue) was used to stain the cell nucleus.
4. Blank control: Raji (blue). Primary Antibody: Rabbit Anti-CCR7 antibody (TMAB-00326), Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit Igg (orange),used under the same conditions); Secondary Antibody: Goat anti-rabbit IgG-Pe (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min). Primary antibody (TMAB-00326, 1 μg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice.
5. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (CCR7) polyclonal Antibody, Unconjugated (TMAB-00326) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
6. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (CCR7) polyclonal Antibody, Unconjugated (TMAB-00326) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
7. Paraformaldehyde-fixed, paraffin embedded (RAT lymphoid); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (CCR7) Polyclonal Antibody, Unconjugated (TMAB-00326) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
8. Blank control: THP-1. Primary Antibody (green line): Rabbit Anti-CCR7 antibody (TMAB-00326)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
9. Sample:
Lane 1: Mouse Bone tissue lysates
Lane 2: Mouse Raw264.7 cell lysates
Lane 3: Human Jurkat cell lysates
Lane 4: Human MCF-7 cell lysates
Lane 5: Human THP-1 cell lysates
Primary: Anti-CCR7 (TMAB-00326) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDa

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human CCR7

Human

Beta Chemokine Rec. (CCR),SARS Coronavirus,GPCR