CD4 antibody (AA 385-457)

Catalog No. ABIN671376

Product Details anti-CD4 Antibody

Target: CD4

Binding Specificity: AA 385-457

Reactivity: Human, Mouse, Rat, Chicken, Pig

Host: Rabbit

Clonality: Polyclonal

Conjugate: This CD4 antibody is un-conjugated

Application: Flow Cytometry (FACS), Western Blotting (WB), ELISA, Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p))

Cross-Reactivity: Chicken, Human, Mouse, Pig, Rat

Predicted Reactivity: Dog,Cow,Sheep,Guinea Pig

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from Human CD4 (cytoplasmic domain)

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #029634 (Immunohistochemistry)

by: Confocal Imaging Core, Beth Israel Deaconess Medical Center

No.: #029634

Date: 03/26/2014

Lot Number: 130703

Method validated: Immunohistochemistry

Positive Control: Mouse spleen tissue

Negative Control: Mouse muscle tissue

Notes: Signal was detected in positive control sample and not in negative control sample.

Validation Images

Figure 1: Mouse spleen stained with anti-CD4 (brown) and counterstained with hematoxylin.

Figure 2: Mouse muscle stained with anti-CD4 (brown) and counterstained with hematoxylin.

Figure 3: Mouse spleen stained with isotype control antibody (brown) and counterstained with hematoxylin.

Figure 4: Mouse spleen stained with secondary antibody only (brown) and counterstained with hematoxylin.

Full Methods

Primary Antibody:

• Antibody: CD4 Molecule (CD4)
• Catalog number: ABIN671376
• Lot number: 130703

Secondary Antibody:

• Antibody: Donkey anti-Rabbit IgG Antibody (Biotinylated)

Full Protocol:

• Immunohistochemistry was performed manually.
• Sections were deparaffinized and rehydrated.
• Sections were heated to 98°C for 10 min in 0.1M Borate buffer with 1 mM NaCl and 1 mM EDTA pH 8.0 for antigen retrieval.
• Sections were incubated in 3% hydrogen peroxide for 10 min at room temperature to block endogenous peroxidases.
• Sections were washed x 3 in Tris buffered saline (TBS).
• Sections were blocked in 5 % normal donkey serum for 60 min at room temperature.
• Sections were incubated with primary antibody diluted 1:200 in 5% normal donkey serum in TBS. Incubated at 4°C temperature overnight.
• Sections were washed x 3 in Tris buffered saline.
• Sections were incubated with secondary antibody diluted 1:400 in 5% normal donkey serum in TBS. Incubated for 60 min at room temperature..
• Sections were washed x 3 in Tris buffered saline.
• Sections were incubated with Vectastain ABC kit (Vector Lab PK6100) to enhance signal.
• Sections were incubated with DAB chromogenic substrate (Vector Lab SK-4105) for 60 sec at RT.
• Sections were washed x 3 in Distilled Water.
• Sections were counterstained with hematoxylin for 20 sec.
• Sections were washed x 1 in Distilled Water.
• Sections were dehydrated, mounted and photographed under a Zeiss AxioImager M1 light microscope.

Experimental Notes: - No experimental challenges noted.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

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

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-CD4 antibody (ABIN671376)

Jaime-Ramirez, Yu, Caserta, Yoo, Zhang, Lee, Hofmeister, Lee, Kumar, Pan, Kumar, Baiocchi, Teknos, Pichiorri, Kaur, Old: "Reolysin and Histone Deacetylase Inhibition in the Treatment of Head and Neck Squamous Cell Carcinoma." in: Molecular therapy oncolytics, Vol. 5, pp. 87-96, (2017) (PubMed).

Zhang, Fu, Li, Wang, Zhang, Liu: "Bone morphogenetic protein 7 is associated with the nodal invasion of colon cancer." in: Oncology letters, Vol. 11, Issue 3, pp. 1707-1712, (2016) (PubMed).

Smit, Bester, van den Heever, Schlegel, Botes, Dohmen: "Does prolonged post-mortem cold ischemic harvesting time influence cryopreserved pulmonary homograft tissue integrity?" in: Cell and tissue banking, (2015) (PubMed).

Target Details for CD4

Target: CD4

Alternative Name: CD4 (CD4 Products)

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

Background:

Synonyms: T-cell surface glycoprotein CD4, Cd4, T-cell surface antigen T4/Leu-3, W3/25 antigen

Background: Integral membrane glycoprotein that plays an essential role in the immune response and serves multiple functions in responses against both external and internal offenses. In T-cells, functions primarily as a coreceptor for MHC class II molecule:peptide complex. The antigens presented by class II peptides are derived from extracellular proteins while class I peptides are derived from cytosolic proteins. Interacts simultaneously with the T-cell receptor (TCR) and the MHC class II presented by antigen presenting cells (APCs). In turn, recruits the Src kinase LCK to the vicinity of the TCR-CD3 complex. LCK then initiates different intracellular signaling pathways by phosphorylating various substrates ultimately leading to lymphokine production, motility, adhesion and activation of T-helper cells. In other cells such as macrophages or NK cells, plays a role in differentiation/activation, cytokine expression and cell migration in a TCR/LCK-independent pathway. Participates in the development of T-helper cells in the thymus and triggers the differentiation of monocytes into functional mature macrophages.

Gene ID: 24932

UniProt: P01730

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