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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Immunohistochemistry [2]
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- Product number
- PA5-19136 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PPP2R4 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with bovine and canine based on sequence homology. This antibody is tested in Peptide ELISA: antibody detection limit dilution 32,000.
- Reactivity
- Human, Mouse, Rat
- Host
- Goat
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- -20°C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PPP2R4 using PPP2R4 Polyclonal Antibody (Product # PA5-19136) (0.5 µg/mL) in staining of Human Liver lysate (lane 1), Mouse Liver (lane 2) and Rat Liver (lane 3) (35 µg protein in RIPA buffer). Detected by chemiluminescence.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PPP2R4 Polyclonal Antibody (Product # PA5-19136) and a 38 kDa band corresponding to PPP2R4 was observed across all the tissues and cell line tested. The expression of PPP2R4 is reported to be low in Skeletal Muscle. Whole cell extracts (30 µg) of HeLa (Lane 1), tissue extracts (30 µg lysate) of Mouse Brain (Lane 2), Mouse Liver (Lane 3), Mouse Lung (Lane 4) and Mouse Skeletal Muscle (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.5 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG Heavy Chain, Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PPP2R4 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with PPP2R4 Goat Polyclonal Antibody (Product # PA5-19136) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27012) 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). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PPP2R4 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with PPP2R4 Goat Polyclonal Antibody (Product # PA5-19136) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Goat IgG Heavy Chain Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27012) 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). Panel d represents the merged image showing cytoplasmic and nuclear 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 analysis of PPP2R4 in human testis. Samples were incubated with PPP2R4 polyclonal antibody (Product # PA5-19136). Formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of PPP2R4 in human testis. Samples were incubated with PPP2R4 polyclonal antibody (Product # PA5-19136). Formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval.
