Invitrogen

PIM1 Recombinant Polyclonal Antibody (19HCLC)

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

1 Published Figure

2 References

View all (17) PIM1 antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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PIM1 Antibody in Immunocytochemistry (ICC/IF) — 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.

PIM1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — 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.

PIM1 Antibody in Western Blot (WB) — 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).

PIM1 Antibody in Immunocytochemistry, Proximity Ligation Assay (PLA) (ICC/IF, PLA) — 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.

PIM1 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across K-562 , HEL 92.1.7, THP-1, Jurkat owing to their inherent genetic constitution. Relative expression of Serine/threonine-protein kinase pim-1 was observed in K-562, HEL 92.1.7, THP-1, but not in Jurkat using Anti-PIM1 Recombinant Polyclonal Antibody (19HCLC) (Product # 710504) in Western Blot. {RE}

Product Details

710504

Applications	Tested Dilution	Publications
Western Blot (WB)	1 µg/mL	-
Immunohistochemistry (Paraffin) (IHC (P))	1:50	-
Immunocytochemistry (ICC/IF)	1:100	View 1 publication 1 publication
in situ PLA (PLA)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse
Published species	Not Applicable
Host/Isotype	Rabbit / IgG
Expression System	EXPI293
Class	Recombinant Polyclonal
Type	Antibody
Clone	19HCLC
Immunogen	Peptide corresponding to amino acids 115–130 of human PIM-1 View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS
Contains	0.09% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2532727

Product Specific Information

This antibody is predicted to react with bovine, canine, equine, feline, non-human primate, mouse, rabbit, ovine and porcine based on sequence homology.

Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.

Target Information

PIM1, which belongs to the Serine/Threonine protein kinase family, is thought to play a role in signal transduction in blood cells. The protooncogene PIM1 encodes a protein kinase upregulated in prostate cancer. It may affect the structure or silencing of chromatin by phosphorylating HP1 gamma/CBX3. PIM1 is expressed primarily in cells of the hematopoietic and germ line lineages.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

PIM1 Promotes Survival of Cardiomyocytes by Upregulating c-Kit Protein Expression.

Cells

Ebeid DE,Firouzi F,Esquer CY,Navarrete JM,Wang BJ,Gude NA,Sussman MA

Published figure using PIM1 recombinant polyclonal antibody (Product # 710504) in Immunocytochemistry

Mon Aug 31 00:00:00 EDT 2020

PIM1 Promotes Survival of Cardiomyocytes by Upregulating c-Kit Protein Expression.

Cells

Ebeid DE,Firouzi F,Esquer CY,Navarrete JM,Wang BJ,Gude NA,Sussman MA

Published figure using PIM1 recombinant polyclonal antibody (Product # 710504) in Immunocytochemistry

Mon Aug 31 00:00:00 EDT 2020

Bioinformatics

Protein Aliases: Oncogene PIM1; pim-1 kinase 44 kDa isoform; pim-1 oncogene; pim-1 oncogene (proviral integration site 1); proto-oncogene serine/threonine-protein kinase pim-1; Serine/threonine-protein kinase pim-1

Gene Aliases: PIM; Pim-1; PIM1

UniProt ID: (Human) Q5T7H7, (Mouse) P06803

Entrez Gene ID: (Human) 5292, (Mouse) 18712

Molecular Function: non-receptor serine/threonine protein kinase