CD44 Antibody (8E2F3) - BSA Free

Novus Biologicals | Catalog # NBP1-47386

Novus Biologicals
100% Guarantee

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rabbit (Negative)

Cited:

Human, Mouse, Rat, Canine

Predicted:

Baboon (94%), Canine (93%), Equine (93%). Backed by our 100% Guarantee.

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC, SDS-Page

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 8E2F3

Format

BSA Free
View all CD44 Primary Antibodies »

Product Specifications

Immunogen

Purified recombinant fragment of human CD44 (628-699) expressed in E. coli. [Uniprot: P16070]

Reactivity Notes

Not reactive to rabbit per customer review.

Localization

Membrane; Single-pass type I membrane protein.

Marker

Cell Membrane Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

82 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for CD44 Antibody (8E2F3) - BSA Free

Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of paraffin-embedded human lung cancer tissues using anti-CD44 mAb (green), showing membrane localization. DRAQ5 fluorescent DNA dye (blue).
Immunohistochemistry-Paraffin: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunohistochemistry-Paraffin: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunohistochemistry-Paraffin: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of FFPE human breast carcinoma tissues using CD44 antibody (8E2F3). The signal was developed using DAB based detection and the sections were processed for counterstaining with hematoxylin. The antibody generated mainly a membrane staining representative of CD44 protein.
Flow Cytometry: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Flow Cytometry: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Flow Cytometry: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of HeLa cells using anti-CD44 mAb (right) and negative control (left).
Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-CD44 Antibody [8E2F3] conjugated to DyLight 550 (NBP1-47386R) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Western Blot: CD44 Antibody (8E2F3)BSA Free [NBP1-47386]

Western Blot: CD44 Antibody (8E2F3)BSA Free [NBP1-47386]

CD44-Antibody-8E2F3-BSA-Free-Western-Blot-NBP1-47386-img0021.jpg
Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with CD44 Antibody [8E2F3] conjugated to Alexa Fluor 647 (NBP1-47386AF647) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Flow Cytometry: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Flow Cytometry: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Flow Cytometry: CD44 Antibody (8E2F3) [NBP1-47386] - An intracellular stain was performed on HeLa cells with NBP1-47386AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.
Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of methanol-fixed A431 (A), HeLa (B), PANC-1 (C) and EC (D) cells using anti-CD44 mAb (green), showing membrane localization. DRAQ5 fluorescent DNA dye (blue).
Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/ Immunofluorescence: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunocytochemistry/Immunofluorescence: CD44 Antibody (8E2F3) [NBP1-47386] - Analysis of PANC-1 cells using anti-CD44 mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). DRAQ5 fluorescent DNA dye (blue).
Flow (Intracellular): CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Flow (Intracellular): CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Flow (Intracellular): CD44 Antibody (8E2F3) [NBP1-47386] - An intracellular stain was performed on HeLa cells with NBP1-47386AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.
Immunoprecipitation: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunoprecipitation: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunoprecipitation: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386] - HAP1 lysates were prepared and immunoprecipitation was performed using 1.0 ug of the CD44 Antibody (NBP1-47386) pre-coupled to either protein G or protein A Sepharose beads. Ability of the antibodies to capture CD44 antigen was first assessed by comparing the level of CD44 antigen from the starting material (SM) to its level remaining in the unbound fractions (UB). Anti-CD44 antigen at 1/2000 was used for each immunoblot. Immunoprecipitate for CD44 Antibody (NBP1-47386) that showed depleted CD44 antigen in the UB can be seen. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).
CD44 Antibody (8E2F3) - BSA Free

CD44 (8E2F3) in A431 Human Cell Line -

CD44 (8E2F3) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti- CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to Biotin (Catalog # NBP1-47386B) at 2 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
CD44 Antibody (8E2F3) - BSA Free

CD44 (8E2F3) in A431 Human Cell Line -

CD44 (8E2F3) was detected in immersion fixed A431 human skin carcinoma cell line using Mouse anti-CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to DyLight 550 (Catalog # NBP1-47386R) (red) at 5 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
CD44 Antibody (8E2F3) - BSA Free

CD44 (8E2F3) in NIH-3T3 Mouse Cell Line.

CD44 (8E2F3) was detected in immersion fixed NIH3T3 Mouse fibroblast cell line using Mouse anti- CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to DyLight 550 (Catalog # NBP1-47386R) (red) at 2 µg/mL overnight at 4C. Cells were counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
CD44 Antibody (8E2F3) - BSA Free

Detection of CD44 (8E2F3) in U-251 MG Human Cell Line by Flow Cytometry.

U-251 MG human glioblastoma cell line was stained with Mouse anti-CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to Alexa Fluor® 647 (Catalog # NBP1-47386AF647, blue histogram) or matched control antibody (orange histogram).
CD44 Antibody (8E2F3) - BSA Free

Detection of CD44 (8E2F3) in A431 Human Cell Line by Flow Cytometry.

An intracellular stain was performed on A431 human skin carcinoma cell line using Mouse anti-CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to DyLight 550 (Catalog # NBP1-47386R, blue histogram) or matched control antibody (orange histogram) at 2.5 µg/mL for 30 minutes at RT.
CD44 Antibody (8E2F3) - BSA Free

CD44 (8E2F3) in U-2 OS Human Cell Line.

CD44 (8E2F3) was detected in immersion fixed U-2 OS human osteosarcoma cell line using Mouse anti-CD44 (8E2F3) Protein-G purified Monoclonal Antibody conjugated to Biotin (Catalog # NBP1-47386B) at 5 µg/mL overnight at 4C. Cells were stained using Streptavidin conjugated to DyLight 550 (red) and counterstained with DAPI (blue). Cells were imaged using a 100X objective and digitally deconvolved.
CD44 Antibody (8E2F3) - BSA Free Detection of CD44 by Immunoprecipitation.

Detection of CD44 by Immunoprecipitation.

MDA‑MB‑231 human breast cancer cell line lysates were prepared and immunoprecipitation was performed using 2.0 μg of CD44 Antibody (8E2F3) - BSA Free (Catalog # NBP1-47386) pre-coupled to Dynabeads Protein G. Immunoprecipitated CD44 was detected in Western Blot with a rabbit CD44 antibody used at 1/3000. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).
CD44 Antibody (8E2F3) - BSA Free CD44 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

CD44 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

MDA‑MB‑231 human breast cancer parental cell line WT and CD44 MDA-MB-231 KO cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with CD44 Antibody (8E2F3) - BSA Free (Catalog # NBP1-47386) followed by incubation with an Alexa-fluor 555 conjugated secondary antibody (upper panel). DAPI-only counterstained cells shown on a lower panel. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Primary antibody dilution used: 1/1000. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).
CD44 Antibody (8E2F3) - BSA Free Immunohistochemistry-Paraffin: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Immunohistochemistry-Paraffin: CD44 Antibody (8E2F3) - BSA Free [NBP1-47386]

Analysis of a FFPE tissue section of human skin using 1:200 dilution of CD44 (8E2F3) antibody. The staining was developed using HRP labeled anti-mouse secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.

Applications for CD44 Antibody (8E2F3) - BSA Free

Application
Recommended Usage

ELISA

1:10000

Flow Cytometry

1:200-1:400

Immunocytochemistry/ Immunofluorescence

1:200-1:1000

Immunohistochemistry

1:200-1:1000

Immunohistochemistry-Paraffin

1:200-1:1000

Knockout Validated

Knockout validated from YCharOS Inc. (YCharOS.com)

Reviewed Applications

Read 1 review rated 5 using NBP1-47386 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Ammonium sulfate precipitation

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: CD44

CD44, also known as IN, LHRMIC4, CDW44, HCELL, is a cell-surface glycoprotein involved in cell-cell interactions, cell adhesion and migration. It is a receptor for hyaluronic acid (HA) and can also interact with other ligands, such as osteopontin, collagens, and matrix metalloproteinases (MMPs). This protein participates in a wide variety of cellular functions including lymphocyte activation, recirculation and homing, hematopoiesis, and tumor metastasis. Transcripts for this gene undergo complex alternative splicing that results in many functionally distinct isoforms, however, the full length nature of some of these variants has not been determined. Alternative splicing is the basis for the structural and functional diversity of this protein, and may be related to tumor metastasis.

Alternate Names

CD44, ECMR-III, HCAM, HCELL, LHR, MDU2, MDU3, MIC4, MUTCH-I, Pgp1

Entrez Gene IDs

960 (Human)

Gene Symbol

CD44

UniProt

P16070

Additional CD44 Products

Product Documents for CD44 Antibody (8E2F3) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for CD44 Antibody (8E2F3) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for CD44 Antibody (8E2F3) - BSA Free

Customer Reviews for CD44 Antibody (8E2F3) - BSA Free (1)

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Customer Images

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  • Name: Corey.
    Application: Western Blot
    Sample Tested: Human Brain Tissue
    Species: Human
    Verified Customer | Posted 07/12/2013
    Western blot analysis of CD44 in brain tissue
    CD44 Antibody (8E2F3) - BSA Free NBP1-47386

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Protocols

View specific protocols for CD44 Antibody (8E2F3) - BSA Free (NBP1-47386):

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

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FAQs

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