CD4 antibody (CD4 Molecule) (AA 1-390) (FITC)

Catalog No. ABIN2104933

Product Details anti-CD4 Antibody

Target:

CD4

Binding Specificity: AA 1-390

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD4 antibody is conjugated to FITC

Application: Flow Cytometry (FACS)

Specificity: Human CD4

Characteristics: This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified, recombinant Human CD4 (rh CD4, ABIN2002782 NP_000607.1, Met1-Trp390, and conjugated with FITC under optimum conditions, the unreacted FITC was removed.

Immunogen: Recombinant Human CD4 protein (ABIN2002782)

Clone: 08

Isotype: IgG1

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Independent Validation (FACS)

Validation #029847 (Flow Cytometry)

by: Flow Cytometry & Cell Separation Facility, Purdue University

No.: #029847

Date: 08/31/2015

Lot Number: HG08FE1721

Method validated: Flow Cytometry

Positive Control: peripheral blood mononuclear cells

Negative Control: peripheral blood mononuclear cells

Notes: This antibody performed well and is comparable to anti-CD4-FITC from BD Pharmingen (see data below)

Validation Images

Figure 1: Histogram of peripheral blood lymphocytes stained with anti-CD4-FITC (red), unstained cells are shown in blue. Percentage of CD4+ cells is 52.7%, median fluorescence intensity is 3797.

Figure 2: Histogram of peripheral blood lymphocytes stained with anti-CD4-FITC from BD Pharmingen (red), unstained cells are shown in blue. Percentage of CD4+ cells is 52.5%, median fluorescence intensity is 4901.

Figure 3: Dot plot of peripheral blood lymphocytes stained with anti-CD4-FITC (x-axis) and anti-CD3-PE (y-axis).

Full Methods

Primary Antibody:

• Antigen: human CD4 (CD4)
• Catalog number: ABIN2104933
• Lot number: HG08FE1721
• Dilution: 1 µl in 100 µL 1X PBS containing 0.5% BSA

Full Protocol:

• 1. 10mls of blood (collected in EDTA) was diluted to 20mls with PBS + 0.5% BSA
• 2. 10mls of diluted blood was overlaid onto 5mls of ficoll.
• 3. Spin for 20 minutes at 1500 rpm with break off.
• 4. The buffy coat layer was collected and wash with PBS/BSA.
• 5. Residual red blood cells were lysed by resuspending the cells in 1ml ammonium chloride for 5-10 minutes at room temperature.
• 6. The cells were washed with PBS/BSA, counted and resuspended at 10x10^6 cells/ml.
• 7. 100ul of cells (1x10^6) were added to their respective tubes.
• 8. Add 5µl of anti-CD4-FITC (Sino Biological) or 20µl of anti-CD4-FITC (BD Pharmingen) to the appropriate tubes and incubate for 20 minutes on ice.
• 9. Add 1ml PBS/BSA. Spin the cells down and resuspend in 0.5ml PBS/BSA.
• 10. Analyze

Handling

Format: Liquid

Concentration: 0.1 mg/mL

Buffer: Aqueous solution containing 0.5 % BSA and 0.09 % sodium azide

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Protected from prolonged exposure to light. Do not freeze ! Sodium azide is toxic to cells and should be disposed of properly. Flush with large volumes of water during disposal

Storage: 4 °C

Storage Comment: This antibody is stable for 12 months from date of receipt when stored at 2°C-8°C. Protected from prolonged exposure to light. Do not freeze !
Sodium azide is toxic to cells and should be disposed of properly. Flush with large volumes of water during disposal.

Expiry Date: 12 months

Target Details for CD4

Target:

CD4

Alternative Name: CD4 (CD4 Antibody Abstract)

Synonyms: cd4, L3T4, Ly-4, CD4mut, W3/25, p55, CD4 molecule, carcinoembryonic antigen-related cell adhesion molecule 5, T-cell surface glycoprotein CD4, CD4 antigen, Cd4 molecule, CD4, LOC100125525, cd4, Cd4

Pathways: TCR Signaling, Maintenance of Protein Location, CXCR4-mediated Signaling Events