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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
- Other assay [1]
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- Product number
- PA5-34666 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Moesin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: NT2D1, GL261, C8D30, NIH-3T3, BCL-1, Raw264.7, C2C12. Predicted reactivity: Mouse (98%), Rat (97%), Pig (98%), Rhesus Monkey (100%), Bovine (99%). IHC notes, Requires antigen retrieval using heat mediated 10mM Citrate buffer (pH6.0) or Tris-EDTA buffer (pH8.0) Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.21 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Moesin Polyclonal Antibody (Product # PA5-34666). Sample (30 µg of whole cell lysate). Lane A: NT2D1. 7.5% SDS PAGE. Moesin Polyclonal Antibody (Product # PA5-34666) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Moesin Polyclonal Antibody detects MSN protein by Western blot analysis. A. 30 µg GL261 whole cell lysate/extract. B. 30 µg C8D30 whole cell lysate/extract. C. 30 µg NIH-3T3 whole cell lysate/extract. D. 30 µg BCL-1 whole cell lysate/extract. E. 30 µg Raw264.7 whole cell lysate/extract. F. 30 µg C2C12 whole cell lysate/extract.7.5 % SDS-PAGE. Moesin Polyclonal Antibody (Product # PA5-34666) dilution: 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Moesin was achieved by transfecting A-431 cells with Moesin specific siRNAs (Silencer® select Product # s8986, s8984). Western blot analysis (Fig. a) was performed using whole cell extracts from the Moesin knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Moesin Polyclonal Antibody (Product # PA5-34666, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/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 Moesin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of A549 (Lane 1), A-431 (Lane 2), Jurkat (Lane 3), COS-7 (Lane 4), C2C12 (Lane 5), NIH/3T3 (Lane 6) and Mouse Spleen (Lane 7). The blot was probed with Anti-Moesin Polyclonal Antibody (Product # PA5-34666, 1:2500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 68kDa band corresponding to Moesin was observed across all cell lines and tissue lysate tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Moesin in methanol-fixed A431 cells using a Moesin polyclonal antibody (Product # PA5-34666) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (red) at a 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Moesin was performed using 70% confluent log phase A549 cells. 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 Moesin Polyclonal Antibody (Product # PA5-34666) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of Moesin was performed in paraffin-embedded rat spleen tissue using Moesin Polyclonal Antibody (Product # PA5-34666) at a dilution of 1:500.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of Moesin was performed in A431 whole cell extracts using 5 µg of Moesin Polyclonal Antibody (Product # PA5-34666). Samples were transferred to a membrane and probed with Moesin Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.
