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<strong>Figure 1 Western Blot Validation with Human Recombinant Protein</strong><br>Loading: 5ng (Lane 1), 25ng (Lane 2) and 100ng (Lane 3) of human CD4 recombinant protein.Antibodies: CD4 PM-5201 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.Observed at around 27kD.
<strong>Figure 2 Western Blot Validation in Human Thymus Tissue Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: CD4 PM-5201 (Lane 1: 0.5 μg/mL, Lane 2: 1 μg/mL and Lane 3: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.
<strong>Figure 3 Western Blot Validation in Human Jurkat Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: CD4 PM-5201 (Lane 1: 1 μg/mL, Lane 2: 2 μg/mL and Lane 3: 4 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.
<strong>Figure 4 Western Blot Validation in Human, Mouse and Rat Tissue Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: CD4 PM-5201 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.
<strong>Figure 5 Western Blot Validation in Human Cell Lines </strong><br>Loading: 15 μg of lysates per lane.Antibodies: CD4 PM-5201 (4 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.
<strong>Figure 6 Western Blot Validation in Mouse Spleen Tissue Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: CD4 PM-5201 (Lane 1: 1 μg/mL, Lane 2: 2 μg/mL and Lane 3: 4 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.
<strong>Figure 7 Western Blot Validation in Rat Lung Tissue Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: CD4 PM-5201 (Lane 1: 0.5 μg/mL, Lane 2: 1 μg/mL and Lane 3: 2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.
<strong>Figure 8 Immunohistochemistry Validation of CD4 in Human Thymus Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded human thymus tissue using anti-CD4 antibody (PM-5201) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-mouse IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
Figure 1 Western Blot Validation with Human Recombinant Protein
Loading: 5ng (Lane 1), 25ng (Lane 2) and 100ng (Lane 3) of human CD4 recombinant protein.Antibodies: CD4 PM-5201 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.Observed at around 27kD.
Trial Size

CD4 Antibody [9H5A8]

CATALOG NUMBER: PM-5201

Clonality:
Monoclonal
Tested Applications:
ELISA, IHC-P, WB
Host Species:
Mouse
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Mouse
Species Reactivity:
Human, Mouse, Rat
Immunogen:
Mouse monoclonal CD4 antibody was raised against a 193 amino acid recombinant protein from near the amino terminus of human CD4.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB
Application Note:
WB: 0.5-4 μg/mL; IHC: 5 μg/mL.
Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1314 - Human Thymus Tissue Lysate
Positive Control 2:
Cat. No. 1205 - Jurkat Cell Lysate
Predicted Molecular Weight:
Predicted: 51kD
Observed: 51-57 kD (Post-modyfication: 2-4 N-linked glycosylations)
Validation:

Recombinant Protein Test (Figure 1): Anti-CD4 antibodies (PM-5201) detected human CD-4 recombinant protein at 5ng.

Isoforms:
Human CD4 has one isoform (458aa, 51kD). Mouse CD4 has two isoforms, including isoform 1 (457aa, 51kD) and isoform 2 (217aa, 24.5kD). Rat CD4 has one isoform (457aa, 51kD). PM-5201 can detect human, mouse and rat.
Purification:
CD4 Monoclonal Antibody is immunoaffinity chromotography purified IgG.
Clonality:
Monoclonal
Isotype:
IgG1
physical-state:
Liquid
Buffer:
CD4 Monoclonal Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
CD4 monoclonal antibody can be stored at -20°C, stable for one year.
Ncbi Official Symbol:
CD4
Additional Names:
CD4 Antibody [9H5A8] : CD4mut
Protein Accession Number:
NP_000607
Protein Gi Number:
10835167
Ncbi Gene Id Number:
920
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
CD4 Monoclonal Antibody: CD4 is a member of the immunoglobulin superfamily and is implicated as associative recognition element in MHC (Major Histocompatibility Complex) class II-restricted immune response. On T-lymphocytes, it defines the helper/inducer subset. The mature 55 kd CD4 protein consists of a 372 amino acid extracellular segment composed of four tandem immunoglobulin-like VJ regions. The CD4 molecule is a major receptor for human immunodeficiency virus (HIV), binding directly to the envelope glycoprotein gp120 on HIV, with the co-receptors being CCR5 or CXCR4. It has been shown that the V-like domains are critical for binding with HIV envelope gp120.
Background Reference 1:
Bowers et al. Int. J. Biochem. Cell Biol. 1997; 29 :871-5.
Background Reference 2:
Arthos et al. Cell 1989; 57:469-81.
Citation 1:
Hood et al. Cytolytic nanoparticles attenuate HIV-1 infectivity. Antivir Ther. 2013;18(1):95-103. PMID: 22954649

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

PM-5201

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
Shipping Info

$65 Standard Overnight flat rate for as many products shipped within the United States.

$35 Dry Ice Shipment fee may be required on some items.

Note: Online orders only accepted for shipment within the USA. Click here for the list of international distributors.

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