BM2 (Myeloid Marker) Antibody Staining Protocol for Immunohistochemistry

 

Description: NCL-BM2 is reactive with a cytoplasmic antigen expressed in human granulocytes. Markers of myeloid cells are useful in the identification of different levels of cellular differentiation. NCL-BM1 and NCL-BM2 are reported to react with early precursor and mature forms of human myeloid cells.

 

Primary Antibody

Name: BM2 (Myeloid Marker) Antibody

Clone: BM-2, Mouse Anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-BM2

Dilution: 1:25 - 1:50 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60min/room temperature


Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
Staining Pattern: Cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Blood or bone marrow
Fixation: Formalin-fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Blood or bone marrow
Negative Control: Omit primary antibody, isotype control, absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary

Notes:

There is no need to perform antigen retrieval on acetone fixed frozen sections

 

References: