bs-0647R[Primary Antibody]

Rabbit  Anti-CD4 Polyclonal Antibody

DATASHEET

Host:Rabbit

Target Protein:CD4

IR:Immunogen Range:385-457/457

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:920

Swiss Prot:P01730

Source:KLH conjugated synthetic peptide derived from human CD4:385-457/457 

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:This gene encodes a membrane glycoprotein of T lymphocytes that interacts with major histocompatibility complex class II antigenes and is also a receptor for the human immunodeficiency virus. This gene is expressed not only in T lymphocytes, but also in B cells, macrophages, and granulocytes. It is also expressed in specific regions of the brain. The protein functions to initiate or augment the early phase of T-cell activation, and may function as an important mediator of indirect neuronal damage in infectious and immune-mediated diseases of the central nervous system. Multiple alternatively spliced transcript variants encoding different isoforms have been identified in this gene. [provided by RefSeq, Aug 2010].

Size:50ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:500-1000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(1μg/Test)
IF(1:100-500)

Cross Reactive Species:Human
Mouse
Rat
Dog
Pig
Cow
Sheep
Guinea Pig
MACFA.

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

VALIDATION IMAGES

Sample: 
HL60 Cell Lysate at 40 ug
Primary: Anti- CD4 (bs-0647R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 55kD

U937 Cell Lysate at 40 ug
Primary: Anti- CD4 (bs-0647R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 55kD

Sample: 
Lane 1: U937 (Human) Cell Lysate at 30 ug
Lane 2: HL-60 (Human) Cell Lysate at 30 ug
Lane 3: Jurkat (Human) Cell Lysate at 30 ug
Primary: Anti-CD4 (bs-0647R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 55 kD
Observed band size: 55 kD

Sample: 
Spleen (Mouse) Lysate at 40 ug
Primary: Anti- CD4 (bs-0647R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 55 kD

Sample: 
Jurkat(Human) Cell Lysate at 30 ug
Primary: Anti- CD4 (bs-0647R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 55 kD

Sample: 
HUVEC(Human) Cell Lysate at 30 ug
Primary: Anti- CD4 (bs-0647R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 55 kD

Sample: 
Jurkat(Human) Cell Lysate at 30 ug
Primary: Anti- CD4 (bs-0647R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 53 kD

Sample: 
Spleen (Mouse) Lysate at 40 ug
Primary: Anti- CD4 (bs-0647R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 55kD

Sample: 
Lymph node(Mouse) Lysate at 40 ug
Primary: Anti- CD4 (bs-0647R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kD
Observed band size: 55kD

Paraformaldehyde-fixed, paraffin embedded (human tonsil); 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 (CD4) Polyclonal Antibody, Unconjugated (bs-0647R) at 1:4000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); 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 (CD4) Polyclonal Antibody, Unconjugated (bs-0647R) at 1:4000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Image was kindly submitted by Dr.David M Burmeister from US Army Institute of Surgical Research. Pig lymph nodes stained with Rabbit Anti-CD4 Polyclonal Antibody(bs-bs-0647R)at 1:300 for one hour at room temperature.

Blank control:HL-60. 
Primary Antibody (green line): Rabbit Anti-CD4 antibody (bs-0647R-APC) 
Dilution: 2μg /10^6 cells; 
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were 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. Acquisition of 20,000 events was performed.

Blank control:Mouse spleen. 
Primary Antibody (green line): Rabbit Anti-CD4 antibody (bs-0647R-APC) 
Dilution: 2μg /10^6 cells; 
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were 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. Acquisition of 20,000 events was performed.

Blank control:Mouse spleen. 
Primary Antibody (green line): Rabbit Anti-CD4 antibody (bs-0647R-PE) 
Dilution: 2μg /10^6 cells; 
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were 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. Acquisition of 20,000 events was performed.

Blank control: Mouse splen. 
Primary Antibody (green line): Rabbit Anti-CD4 antibody (bs-0647R-FITC) 
Dilution: 3μg /10^6 cells; 
Isotype Control Antibody (orange line): Rabbit IgG .
Dilution: 3μg /test. 
Protocol
The cells 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. Acquisition of 20,000 events was performed.

Blank control (Black line): Molt4 (Black). Primary Antibody (green line): Rabbit Anti-CD4 antibody (bs-0647R-PE) Dilution: 3μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG-PE. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then were 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. Acquisition of 20,000 events was performed.

Blank control (blue line): HL60 (fixed with 2% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature). 
Primary Antibody (green line): Rabbit Anti-CD4 antibody (bs-0647R),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.