PCM1 - Antibodies - The Human Protein Atlas


	Antibody HPA023370	Antibody HPA023374	Antibody CAB058695

ANTIBODY INFORMATION
Provider	Atlas Antibodies Sigma-Aldrich	Atlas Antibodies Sigma-Aldrich	Atlas Antibodies Sigma-Aldrich
Product name	HPA023370	HPA023374	AMAb90565
Host species	Rabbit	Rabbit	Mouse
Clonalityⁱ The antibodies are designated mAB for monoclonal and pAb for polyclonal.	pAb	pAb	mAb
Concentration	0.49 mg/ml	0.6225 mg/ml	Not known
Purity	Affinity purified using the PrEST-antigen as affinity ligand	Affinity purified using the PrEST-antigen as affinity ligand	Protein A/G
Released in versionⁱ The release of the Human Protein Atlas in which the antibody was first published.	5.0	5.0	11.0
Referencesⁱ References to publications in which the antibody has been used.	9	10	1
Proper citation	Atlas Antibodies Cat#HPA023370, RRID:AB_1855072	Atlas Antibodies Cat#HPA023374, RRID:AB_1855073	Atlas Antibodies Cat#AMAb90565, RRID:AB_2665588
Validation summaryⁱ All assays through which the antibody has been validated. Assays&annotation provide a detailed description of the different assays. The pie-charts indicate degree of validation.	ICC IHC N/A WB PA	ICC IHC WB PA	N/A ICC IHC N/A WB N/A PA
IMMUNOCYTOCHEMISTRYⁱ Immunocytochemistry is used to validate the antibody staining and for assessing and validating the protein expression pattern in selected human cell lines. Read more
Validationⁱ Results of validation by standard or enhanced validation. Standard validation is based on concordance with available experimental gene/protein characterization data in the UniProtKB/Swiss-Prot database. Standard validation results in scores Supported, Approved or Uncertain. Enhanced validation is performed using either siRNA knockdown, tagged GFP cell lines or independent antibodies. For the siRNA validation the decrease in antibody-based staining intensity upon target protein downregulation is evaluated. For the GFP validation the signal overlap between the antibody staining and the GFP-tagged protein is evaluated. For the independent antibodies validation the evaluation is based on comparison of the staining of two (or more) independent antibodies directed towards independent epitopes on the protein. For all cases except the siRNA validation, an image representative of the antibody staining pattern is shown. For the siRNA validation, a box plot of the results is shown. Read more	Supportedⁱ Immunocytochemistry is used for validating antibody reliability by assessing staining pattern in cell lines. Validation scores include Enhanced, Supported, Approved and Uncertain. Read more The subcellular location is supported by literature. Immunofluorescent staining of human cell line U-251MG shows localization to nucleoplasm, nuclear membrane, cytosol & centriolar satellites.	Supportedⁱ Immunocytochemistry is used for validating antibody reliability by assessing staining pattern in cell lines. Validation scores include Enhanced, Supported, Approved and Uncertain. Read more The subcellular location is supported by literature. Immunofluorescent staining of human cell line U-251MG shows localization to cytosol & centriolar satellites.	N/A

Antibody dilution	Human assay: A-431 fixed with PFA, dilution: 1:200 Human assay: U-251MG fixed with PFA, dilution: 1:200 Human assay: U2OS fixed with PFA, dilution: 1:200	Human assay: A-431 fixed with PFA, dilution: 1:400 Human assay: U-251MG fixed with PFA, dilution: 1:400 Human assay: U2OS fixed with PFA, dilution: 1:400
IMMUNOHISTOCHEMISTRYⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain. Read more
Validationⁱ Results of validation by standard or enhanced validation based on assessment of antibody performance in 44 normal tissues. Standard validation results in scores Supported, Approved or Uncertain. An image representative of the antibody staining pattern is shown. Enhanced validation results in the score Enhanced and includes two methods: Orthogonal validation and Independent antibody validation. For orthogonal validation, representative images of high and low expression are shown. For independent antibody validation, four images of each independent antibody are displayed. Read more	Enhanced - Independent antibodies Protein distribution across 45 tissues similar between the independent antibodies HPA023370 and HPA023374. Kidney Liver Ovary Testis	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 45 tissues. HIGH EXPRESSION Testis RNA expression: 154.2 nTPM LOW EXPRESSION Duodenum RNA expression: 21.8 nTPM Enhanced - Independent antibodies Protein distribution across 45 tissues similar between the independent antibodies HPA023370 and HPA023374. Kidney Liver Ovary Testis	Supportedⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain. Read more Immunohistochemical staining of human ovary shows strong positivity in oocytes. Ovary

Retrievalⁱ Antigen retrieval is a method used to restore/retrieve the epitope (antibody bidning region) of the target protein, cross-linked, and thus masked, during tissue preserving fixative treatment of the tissues. Read more	HIER pH6	HIER pH6	HIER pH6
Antibody dilution	1:2000	1:300	1:175
Literature conformityⁱ Conformance of the expression pattern with available gene/protein characterization data in scientific literature and data from bioinformatic predictions. UniProt is used as the main source of gene/protein characterization data and when relevant, available publications and other sources of information are researched in depth. Extensive or sufficient gene/protein data requires that there is evidence of existence on a protein level and that a substantial quantity of published experimental data is available from literature and public databases. Limited protein/gene characterization data does not require evidence of existence on a protein level and refers to genes for which only bioinformatic predictions and scarce published experimental data is available.	Consistent with extensive gene/protein characterization data.	Consistent with extensive gene/protein characterization data.	Consistent with extensive gene/protein characterization data.
RNA consistencyⁱ Consistency between immunohistochemistry data and consensus RNA levels is divided into five different categories: i) High consistency, ii) Medium consistency, iii) Low consistency, iv) Very low consistency, and v) Cannot be evaluated.	Medium consistency between antibody staining and RNA expression data.	Medium consistency between antibody staining and RNA expression data.	Medium consistency between antibody staining and RNA expression data.
WESTERN BLOTⁱ A Western blot analysis is performed on a panel of human tissues and cell lines to evaluate antibody specificity. For antibodies with unreliable result a revalidation using an over-expression lysate is performed. Read more
Validationⁱ Western Blot is used for quality control of the polyclonal antibodies generated in the project. After purification, the antibodies are used to detect bands in a setup of lysate and different tissues. The result is then scored Enhanced, Supported, Approved, or Uncertain. Enhanced validation includes five different methods: Genetic validation, Recombinant expression validation, Independent antibody validation, Orthogonal validation and Capture MS validation. Read more	Uncertainⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Weak band of predicted size but with additional bands of higher intensity also present. Analysis performed using a standard panel of samples.	Supportedⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Band of predicted size in kDa (+/-20%) with additional bands present. Analysis performed using a standard panel of samples. 230 130 95 72 56 36 28 17 11	Uncertainⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Weak band of predicted size but with additional bands of higher intensity also present. Analysis performed using a standard panel of samples.

Antibody dilution	1:250	1:250	1:500
PROTEIN ARRAY
Validationⁱ A protein array containing 384 different antigens including the antibody target is used to analyse antibody specificity. Depending on the array interaction profile the antibody is scored as Supported, Approved, or Uncertain. Read more	Approved Pass with quality comment low specificity (binding to 1-2 antigens >15% and <40%). Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.	Supported Pass with single peak corresponding to interaction only with its own antigen. Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.	N/A

Antibody dilution	1:120000	1:3000
RELEVANT PUBLICATIONS
	Dynamic DNA methylation orchestrates cardiomyocyte development, maturation and disease Gilsbach R et al Nat Commun 2014;5:5288 Application: FC, IHC	Identification of cardiomyocyte nuclei and assessment of ploidy for the analysis of cell turnover Bergmann O et al Exp Cell Res 2011;317(2):188-94	Centriolar Satellites Control GABARAP Ubiquitination and GABARAP-Mediated Autophagy Joachim J et al Curr Biol 2017;27(14):2123-2136.e7 Application: WB
	RBM14 prevents assembly of centriolar protein complexes and maintains mitotic spindle integrity Shiratsuchi G et al EMBO J 2015;34(1):97-114 Application: ICC-IF	ARL13B, PDE6D, and CEP164 form a functional network for INPP5E ciliary targeting Humbert MC et al Proc Natl Acad Sci U S A 2012;109(48):19691-6
	Cardiac Myocyte De Novo DNA Methyltransferases 3a/3b Are Dispensable for Cardiac Function and Remodeling after Chronic Pressure Overload in Mice Nührenberg TG et al PLoS One 2015;10(6):e0131019 Application: IHC	BBS mutations modify phenotypic expression of CEP290-related ciliopathies Zhang Y et al Hum Mol Genet 2014;23(1):40-51 Application: ICC-IF, WB
	Epicardial FSTL1 reconstitution regenerates the adult mammalian heart Wei K et al Nature 2015;525(7570):479-85 Application: ICC-IF, IHC	Single cardiomyocyte nuclear transcriptomes reveal a lincRNA-regulated de-differentiation and cell cycle stress-response in vivo See K et al Nat Commun 2017;8(1):225 Application: ICC-IF
	Alterations in sarcomere function modify the hyperplastic to hypertrophic transition phase of mammalian cardiomyocyte development Nixon BR et al JCI Insight 2017;2(4):e90656 Application: IHC	A Method for the Acute and Rapid Degradation of Endogenous Proteins Clift D et al Cell 2017;171(7):1692-1706.e18 Application: WB
	NEK7 is required for G1 progression and procentriole formation Gupta A et al Mol Biol Cell 2017;28(15):2123-2134 Application: ICC-IF, WB	DNA methylation signatures follow preformed chromatin compartments in cardiac myocytes Nothjunge S et al Nat Commun 2017;8(1):1667 Application: ChIP
	Hippo pathway deficiency reverses systolic heart failure after infarction Leach JP et al Nature 2017;550(7675):260-264 Application: IHC	Paracrine effect of regulatory T cells promotes cardiomyocyte proliferation during pregnancy and after myocardial infarction Zacchigna S et al Nat Commun 2018;9(1):2432 Application: IHC
	Distinct epigenetic programs regulate cardiac myocyte development and disease in the human heart in vivo Gilsbach R et al Nat Commun 2018;9(1):391 Application: FC	5'-Hydroxymethylcytosine Precedes Loss of CpG Methylation in Enhancers and Genes Undergoing Activation in Cardiomyocyte Maturation Kranzhöfer DK et al PLoS One 2016;11(11):e0166575 Application: FC
	PAN-INTACT enables direct isolation of lineage-specific nuclei from fibrous tissues Bhattacharyya S et al PLoS One 2019;14(4):e0214677 Application: ICC-IF	Dietary protein restriction throughout intrauterine and postnatal life results in potentially beneficial myocardial tissue remodeling in the adult mouse heart Hennig M et al Sci Rep 2019;9(1):15126 Application: ICC-IF
		A liquid-like spindle domain promotes acentrosomal spindle assembly in mammalian oocytes So C et al Science 2019;364(6447): Application: ICC-IF
ANTIGEN INFORMATION
Antigen	Recombinant protein fragment	Recombinant protein fragment	Recombinant protein
Length (aa)	96	95
Antigen sequence	`EEEGVSGASLSSHRSSLVDEHPEDAEFEQKINRLMAAKQKLRQLQDLVAM VQDDDAAQGVISASASNLDDFYPAEEDTKQNSNNTRGNANKTQKDT`	`TIYSEVATLISQNESRPHFLIELFHELQLLNTDYLRQRALYALQDIVSRH ISESHEKGENVKSVNSGTWIASNSELTPSESLATTDDETFEKNFE`
Matching transcripts	PCM1-201 - ENSP00000327077 [100%] PCM1-211 - ENSP00000431099 [100%] PCM1-221 - ENSP00000430521 [100%] PCM1-203 - ENSP00000428131 [85%]	PCM1-201 - ENSP00000327077 [100%] PCM1-211 - ENSP00000431099 [100%] PCM1-214 - ENSP00000429054 [100%] PCM1-221 - ENSP00000430521 [100%]
Matching mouse transcripts	ENSMUSP00000039709 [78%] ENSMUSP00000147887 [78%] ENSMUSP00000155052 [25%] ENSMUSP00000113014 [24%]	ENSMUSP00000039709 [94%] ENSMUSP00000147887 [94%] ENSMUSP00000108838 [25%] ENSMUSP00000027852 [24%]
ANTIGEN VIEWⁱ The protein browser displays the antigen location on the target protein(s) and the features of the target protein. The tabs at the top of the protein view section can be used to switch between the different splice variants to which an antigen has been mapped. At the top of the view, the position of the antigen (identified by the corresponding HPA identifier) is shown as a green bar. A yellow triangle on the bar indicates a <100% sequence identity to the protein target. Below the antigens, the maximum percent sequence identity of the protein to all other proteins from other human genes is displayed, using a sliding window of 10 aa residues (HsID 10) or 50 aa residues (HsID 50). The region with the lowest possible identity is always selected for antigen design, with a maximum identity of 60% allowed for designing a single-target antigen (read more). The curve in blue displays the predicted antigenicity i.e. the tendency for different regions of the protein to generate an immune response, with peak regions being predicted to be more antigenic.The curve shows average values based on a sliding window approach using an in-house propensity scale. (read more). If a signal peptide is predicted by a majority of the signal peptide predictors SPOCTOPUS, SignalP 4.0, and Phobius (turquoise) and/or transmembrane regions (orange) are predicted by MDM, these are displayed. Low complexity regions are shown in yellow and InterPro regions in green. Common (purple) and unique (grey) regions between different splice variants of the gene are also displayed (read more), and at the bottom of the protein view is the protein scale.
PCM1-201 PCM1-203 PCM1-211 PCM1-214 PCM1-221