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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Other assay [1]
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- Product number
- 710504 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PIM1 Recombinant Polyclonal Antibody (19HCLC)
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with bovine, canine, equine, feline, non-human primate, mouse, rabbit, ovine and porcine based on sequence homology.
- Antibody clone number
- 19HCLC
- Concentration
- 0.5 mg/mL
Submitted references PIM1 Promotes Survival of Cardiomyocytes by Upregulating c-Kit Protein Expression.
Ebeid DE, Firouzi F, Esquer CY, Navarrete JM, Wang BJ, Gude NA, Sussman MA
Cells 2020 Aug 31;9(9)
Cells 2020 Aug 31;9(9)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Pim-1 in whole cell extracts of Raji and Jurkat cells (lanes 1-2) using a Pim-1 Recombinant Rabbit Polyclonal Antibody (Product # 710504) at a dilution of 1 µg/mL. Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody followed by chemiluminescence (ECL). Results show a band at ~34kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PIM1 Recombinant Polyclonal Antibody (19HCLC) (Product # 710504) and a 30-45 kDa band corresponding to Serine/threonine-protein kinase pim-1 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of K-562 (Lane 1), HEL 92.1.7 (Lane 2), THP-1 (Lane 3), Jurkat (Lane 4) 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 (1 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescentdetection 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 Serine/threonine-protein kinase pim-1 was performed using 70 confluent log phase K-562 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 PIM1 Recombinant Polyclonal Antibody (19HCLC) (Product # 710504) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (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 withRhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents Jurkat. Panel f 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 Pim-1 showing staining in the cytoplasm of paraffin-embedded mouse testis tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Anti- Pim-1 Antibody (19HCLC) Recombinant Rabbit Polyclonal (Product # 710504) diluted in 3% BSA-PBS at a dilution of 1:50 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 2 PIM1 interacts with c-Kit. ( a ) Immunoblot analysis of a co-immunoprecipitation using whole heart lysates from PIM1 hearts in which PIM1 was pulled down. ( b ) Interaction between PIM1 and c-Kit as determined by proximity ligation assay (top panel) and in the absence of primary antibody (bottom panel). Proximity events are shown as red dots.
