CD4 antibody (AA 385-457)

Catalog No. ABIN671376

This anti-CD4 antibody is a Rabbit Polyclonal antibody detecting CD4 in FACS, WB, ELISA, IHC (fro), IHC (p), IF (cc) and IF (p). Suitable for Human, Mouse, Rat, Chicken and Pig. Independently validated for use in Immunohistochemistry. This Primary Antibody has been cited in 4+ publications.

Quick Overview for CD4 antibody (AA 385-457) (ABIN671376)

Target: CD4

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))

Product Details anti-CD4 Antibody

Binding Specificity: AA 385-457

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

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