14-0718-93

antibody from Invitrogen Antibodies

Targeting: TFRC CD71, p90, TFR1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

14-0718-93

Provider

Invitrogen Antibodies

Product name

CD71 (Transferrin Receptor) Monoclonal Antibody (MRQ-48), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The MRQ-48 monoclonal antibody recognizes human CD71, also known as the transferrin receptor. CD71 exists as a homodimer on the cell surface and is essential for cellular growth. CD71 is expressed on placental syncytiotrophoblasts, endocrine pancreas, hepatocytes, myocytes, basal keratinocytes, spermatocytes, and erythroid precursors. CD71 also is expressed by immature proliferating cells, at low levels on resting mature lymphocytes, and upregulated after antigen or mitogen stimulation of T and B cells. CD71 expression is used to aid in the identification of erythroid leukemia, benign erythroid proliferative disorders, and myelodysplastic syndrome. Applications Reported: This MRQ-48 antibody has been reported for use in immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, microscopy, and immunohistochemical staining. Applications Tested: This MRQ-48 antibody has been tested by immunohistochemistry of formalin-fixed paraffin embedded human tissue using low pH antigen retrieval. This antibody can be used at less than or equal to 1:100. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

MRQ-48

Vial size

100 µL

Storage

4° C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of CD71 was achieved by transfecting HeLa cells with SCRIB specific siRNAs (Silencer® select Product # s725). Western blot analysis (Fig. a) was performed using membrane enriched cell extracts from the CD71 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with CD71 (Transferrin Receptor) Monoclonal Antibody (MRQ-48) (Product # 14-0718-93, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to CD71.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), THP-1 (Lane 2), PC-3 (Lane 3), LNCaP (Lane 4), MCF7 (Lane 5), MDA-MB-231 (Lane 6) and K-562 (Lane 7). The blot was probed with Anti-CD71 (Transferrin Receptor) Monoclonal Antibody (Product # 14-0718-93, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 84 kDa band corresponding to CD71 (Transferrin Receptor) was observed across the cell lines tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CD71 was performed using 70% confluent log phase THP-1 cells differentiated into Macrophage (M0). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CD71(Transferrin Receptor) Mouse Monoclonal Antibody (MRQ-48) (Product # 14-0718-93) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization. Panel e shows untreated cells with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry of formalin-fixed paraffin embedded human placenta tissue using 1:100 Anti-Human CD71 Purified followed by Anti-Mouse Biotin, Streptavidin-HRP, and DAB visualization. Nuclei are counterstained with hematoxylin.