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

ANTIBODY INFORMATION

 
 

Antibody HPA008422

 

Antibody HPA023900

 

Antibody CAB022098

 

Provider

Atlas Antibodies
Sigma-Aldrich
  Atlas Antibodies
Sigma-Aldrich
  Upstate
 

Product name

HPA008422   HPA023900   05-361  

Host species

Rabbit   Rabbit   Mouse  

Clonality

pAb   pAb   mAb  

Purity

Affinity purified using the PrEST-antigen as affinity ligand   Affinity purified using the PrEST-antigen as affinity ligand   Protein A/G  

Other gene match

     

Released in version

4   5   5  

References

1      

Proper citation

Atlas Antibodies Cat#HPA008422, RRID:AB_1854434   Atlas Antibodies Cat#HPA023900, RRID:AB_1854435    

VALIDATION SUMMARY

 
 

Antibody HPA008422

 

Antibody HPA023900

 

Antibody CAB022098

 
Immunocytochemistry

Supported     Supported  
Immunohistochemistry


Validated
 

Validated

Validated
  Supported  
Western blot

Supported   Supported   Supported  
Protein array

Supported   Supported    

IMMUNOCYTOCHEMISTRY

 
 

Antibody HPA008422

 

Antibody HPA023900

 

Antibody CAB022098

 

Standard validation

Supported     Supported  

     

Figure description

Immunofluorescent staining of human cell line A-431 shows localization to cytosol.     Immunofluorescent staining of human cell line U-2 OS shows localization to nucleoplasm & cytosol.  

Antibody dilution

1:38     1:100  

Literature conformity

The subcellular location is supported by literature.     The subcellular location is supported by literature.  

IMMUNOHISTOCHEMISTRY

 
 

Antibody HPA008422

 

Antibody HPA023900

 

Antibody CAB022098

 

Formal validation: Orthogonal

 
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.    
 

Formal validation: Independent

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 HPA023900 across 46 cell lines.
  Distribution of protein expression (antibody staining).
Pearson correlation with HPA008422 across 46 cell lines.
   
 

Standard validation

Supported   Supported   Supported  

     

Figure description

Immunohistochemical staining of human lymph node shows strong cytoplasmic positivity in reaction center cells.   Immunohistochemical staining of human tonsil shows distinct cytoplasmic positivity in most cells.   Immunohistochemical staining of human pancreas shows strong positivity in enocrine and exocrine cells.  

Expression

RNA: detected in 37 tissues
Protein: detected in 58 cell types
  RNA: detected in 37 tissues
Protein: detected in 40 cell types
  RNA: detected in 37 tissues
Protein: detected in 73 cell types
 

Retrieval

HIER pH6   HIER pH6   HIER pH6  

Antibody dilution

1:40   1:15   1:400  

Literature conformity

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

RNA consistency

Mainly consistent with RNA expression data.   Mainly consistent with RNA expression data.   Mainly consistent with RNA expression data.  

WESTERN BLOT

 
 

Antibody HPA008422

 

Antibody HPA023900

 

Antibody CAB022098

 

Standard validation

Supported

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

Analysis performed using a standard panel of samples.
Band of predicted size in kDa (+/-20%) with additional bands 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
  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
  Lane 1: Marker [kDa] 250, 130, 95, 72, 55, 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)

96.7   96.7, 45.6   96.7, 45.6, 7.4, 2.5  

Antibody dilution

1:250   1:250   1:500  

PROTEIN ARRAY

 
 

Antibody HPA008422

 

Antibody HPA023900

 

Antibody CAB022098

 

Standard validation

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   1:500    

ANTIGEN INFORMATION

 
 

Antibody HPA008422

 

Antibody HPA023900

 

Antibody CAB022098

 

Antigen

Recombinant protein fragment   Recombinant protein fragment   Recombinant protein  

Length (aa)

131   135    

Antigen sequence

LLNAAQNTMEPPLTPPSPAGPGLSLGDTALQNLEQLLDGPEAQGSWAELA
ERLGLRSLVDTYRQTTSPSGSLLRSYELAGGDLAGLLEALSDMGLEEGVR
LLRGPETRDKLPSTEVKEDSAYGSQSVEQEA
 
ICNYEGPAKIEVDLVTHSDPPRAHAHSLVGKQCSELGICAVSVGPKDMTA
QFNNLGVLHVTKKNMMGTMIQKLQRQRLRSRPQGLTEAEQRELEQEAKEL
KKVMDLSIVRLRFSAFLRASDGSFSLPLKPVISQP
   

Matching transcripts

NFKB2-001 - ENSP00000189444 [100%]
NFKB2-201 - ENSP00000410256 [100%]
NFKB2-003 - ENSP00000358983 [99%]
  NFKB2-001 - ENSP00000189444 [100%]
NFKB2-003 - ENSP00000358983 [100%]
NFKB2-201 - ENSP00000410256 [100%]
NFKB2-202 - ENSP00000480211 [100%]
   

Other gene match

     
ANTIGEN VIEW
NFKB2-001
NFKB2-003
NFKB2-201
NFKB2-202

RELEVANT PUBLICATIONS

 
 

Antibody HPA008422

 

Antibody HPA023900

 

Antibody CAB022098

 

1.

A roadmap of constitutive NF-κB activity in Hodgkin lymphoma: Dominant roles of p50 and p52 revealed by genome-wide analyses
de Oliveira KA et al
Genome Med 2016;8(1):28
Application: IHC
     
 

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