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Antibodies

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ANTIBODY INFORMATION VALIDATION SUMMARY IMMUNOCYTOCHEMISTRY IMMUNOHISTOCHEMISTRY WESTERN BLOT PROTEIN ARRAY ANTIGEN INFORMATION RELEVANT PUBLICATIONS

ANTIBODY INFORMATION

 
 

Antibody HPA021023

 

Antibody HPA022124

 

Provider

Atlas Antibodies
Sigma-Aldrich
  Atlas Antibodies
Sigma-Aldrich
 

Product name

HPA021023   HPA022124  

Host species

Rabbit   Rabbit  

Clonalityi

The antibodies are designated mAB for monoclonal and pAb for polyclonal.

pAb   pAb  

Purity

Affinity purified using the PrEST-antigen as affinity ligand   Affinity purified using the PrEST-antigen as affinity ligand  

Other gene matchi

Genes, excluding the target gene, to which the antigen sequence of the antibody is >80% identical.

   

Released in versioni

The release of the Human Protein Atlas in which the antibody was first published.

5   5  

Referencesi

References to publications in which the antibody has been used.

   

VALIDATION SUMMARYi

Read more

 
 

Antibody HPA021023

 

Antibody HPA022124

 
Immunohistochemistryi
Immunohistochemistry is used to validate the antibody staining and for assessing and validating the protein expression pattern in normal human tissues. For validation of antibody staining the immunohistochemical staining patterns obtained using tissue microarrays (TMA) containing 44 different normal tissue types (IHC tissue) , as well as, cell microarrays (CMA) containing 44 different widely used and well characterized human cell lines (IHC cells) , are compared to staining patterns from independent antibodies or to RNA expression patterns. For validation of protein expression patterns in normal human tissues the TMA staining patterns are evaluated together with RNA-seq data from internal and external sources and available protein/gene characterization data.

Read more


Validated

Validated
 

Validated

Validated
 
Western bloti
A Western blot analysis is performed on a panel of human tissues and cell lines to evaluate antibody specificity. For antibodies with uncertain result a revalidation using an over-expression lysate is performed.

Read more

Uncertain   Supported  
Protein arrayi
A protein microarray containing 384 different antigens including the antibody target is used for analysis of antibody specificity.

Read more

Supported   Supported  

IMMUNOHISTOCHEMISTRYi

Immunohistochemistry is used to validate the antibody staining and for assessing and validating the protein expression pattern in normal human tissues. For validation of antibody staining the immunohistochemical staining patterns obtained using tissue microarrays (TMA) containing 44 different normal tissue types (IHC tissue) , as well as, cell microarrays (CMA) containing 44 different widely used and well characterized human cell lines (IHC cells) , are compared to staining patterns from independent antibodies or to RNA expression patterns. For validation of protein expression patterns in normal human tissues the TMA staining patterns are evaluated together with RNA-seq data from internal and external sources and available protein/gene characterization data.

Read more

 
 

Antibody HPA021023

 

Antibody HPA022124

 

Formal validation: Orthogonali

Formal validation based on orthogonal methods using cell line data.

Go to Data from the Human Protein Atlas in XML format to download cell line protein expression data and images.

Read more

Validated

Pearson correlation >0.6 for protein and RNA expression in cell lines.
 
Validated

Pearson correlation >0.6 for protein and RNA expression in cell lines.
 

 

 

 
 

 

 

Figure description

Distribution of protein expression (antibody staining) and RNA expression (TPM) across 44 cell lines.   Distribution of protein expression (antibody staining) and RNA expression (TPM) across 44 cell lines.  
 

Formal validation: Independenti

Formal validation based on independent antibodies using cell line data.

Go to Data from the Human Protein Atlas in XML format to download cell line protein expression data and images.

Read more

Validated

Pearson correlation >0.6 for protein expression in cell lines using independent antibodies.
 
Validated

Pearson correlation >0.6 for protein expression in cell lines using independent antibodies.
 

 

 
 

 

Figure description

Distribution of protein expression (antibody staining).
Pearson correlation with HPA022124 across 46 cell lines.
  Distribution of protein expression (antibody staining).
Pearson correlation with HPA021023 across 46 cell lines.
 
 

Formal validation: Independenti

Formal validation based on independent antibodies using tissue data.

Read more

Validated

Spearman correlation >0.6 for protein expression in tissues using independent antibodies.
 
Validated

Spearman correlation >0.6 for protein expression in tissues using independent antibodies.
 

 

 
 

 

Figure descriptioni

Read more

Distribution of protein expression (antibody staining).
Spearman correlation with HPA022124 across 72 cell types.
  Distribution of protein expression (antibody staining).
Spearman correlation with HPA021023 across 72 cell types.
 
 

Standard validationi

For each antibody the observed staining pattern is assigned a validation score based on the conformance with UniProt gene/protein characterization data, as well as, the consistency with RNA expression. The validation score levels are supported, approved, and uncertain.

Read more

Supported   Supported  

   

Figure description

Immunohistochemical staining of human adrenal gland shows strong cytoplasmic and nuclear positivity in cortical cells.   Immunohistochemical staining of human stomach shows distinct cytoplasmic positivity in glandular cells.  

Expression

RNA: detected in 37 tissues
Protein: detected in 30 cell types
  RNA: detected in 37 tissues
Protein: detected in 51 cell types
 

Retrieval

HIER pH6   HIER pH6  

Antibody dilution

1:600   1:200  

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.

Partly consistent with extensive gene/protein characterization data.   Partly consistent with extensive gene/protein characterization data.  

RNA consistencyi

Consistency between immunohistochemistry data and internally generated RNA-seq data is divided into five different categoreies: i) Consistent with RNA expression data, ii) Mainly consistent with RNA expression data, iii) Mainly not consistent with RNA expression data, iv) Not consistent with RNA expression data, and v) No internal RNA expression data available for correlation.

Mainly consistent with RNA expression data.   Mainly consistent with 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

 
 

Antibody HPA021023

 

Antibody HPA022124

 

Standard 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 Supported, Approved, or Uncertain.

Read more

Uncertain

Analysis performed using a standard panel of samples.
Weak band of predicted size but with additional bands of higher intensity also present.
  Supported

Analysis performed using a standard panel of samples.
Band of predicted size in kDa (+/-20%) with additional bands present.
 

   

Figure description

  Lane 1: Marker [kDa] 230, 130, 95, 72, 56, 36, 28, 17, 11
Lane 2: RT4
Lane 3: U-251 MG
Lane 4: Human Plasma
Lane 5: Liver
Lane 6: Tonsil
 

Target mass (kDa)

36.3   36.3  

Antibody dilution

1:250   1:250  

PROTEIN ARRAYi

A protein microarray containing 384 different antigens including the antibody target is used for analysis of antibody specificity.

Read more

 
 

Antibody HPA021023

 

Antibody HPA022124

 

Standard 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

Supported

Pass with single peak corresponding to interaction only with its own antigen.
  Supported

Pass with single peak corresponding to interaction only with its own antigen.
 

   

Figure description

Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.   Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.  

Antibody dilution

1:3000   1:3000  

ANTIGEN INFORMATION

 
 

Antibody HPA021023

 

Antibody HPA022124

 

Antigen

Recombinant protein fragment   Recombinant protein fragment  

Length (aa)

81   83  

Antigen sequence

GCQVVAPSDMMDGRVEAIKEALMAHGLGNRVSVMSYSAKFASCFYGPFRD
AAKSSPAFGDRRCYQLPPGARGLALRAVDRD
 
VLIFGVPSRVPKDERGSAADSEESPAIEAIHLLRKTFPNLLVACDVCLCP
YTSHGHCGLLSENGAFRAEESRQRLAEVALAYA
 

Matching transcripts

ALAD-001 - ENSP00000386284 [100%]
  ALAD-001 - ENSP00000386284 [100%]
 

Other gene matchi

Genes, excluding the target gene, to which the antigen sequence of the antibody is >80% identical.

   
ANTIGEN VIEW
ALAD-001
 

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