Mouse anti-Human IgA (Alpha heavy chain) Secondary Antibody

Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody for Western blot, IHC (P), FACS, ELISA

>> See 2 other secondary antibodies for IGHA1 (alpha heavy chain, immunoglobulin alpha chain, IgA heavy chain)
Synonyms:
alpha heavy chain, immunoglobulin alpha chain, IgA heavy chain
Entrez Gene ID:
UniProt ID:
Details
Host / Isotype: Mouse / IgG1, kappa
Class: Monoclonal
Type: Secondary Antibody
Clone: GA01
Label: Unconjugated
Tested Species Reactivity: Human (Hu)
Immunogen: Purified human alpha heavy chain
Ordering Information
Pierce Mouse anti-Human IgA (Alpha heavy chain) Secondary Antibody
Product #
MA5-11208
Size
500 ul
Price
$185.00
Purchase
Add Mouse anti-Human IgA (Alpha heavy chain) Secondary Antibody to your cart.
Tested Applications Dilution *
Western Blot (WB) 1:00-1:1000
Immunohistochemistry (Paraffin) (IHC (P)) 1:12.5-1:25
Flow Cytometry (FACS) 2 ug/test
ELISA (ELISA) 1 µg/ml
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Form Information
Form: Liquid
Concentration: 0.2mg/ml
Purification: Protein G
Storage Buffer: PBS, pH 7.4, with 0.2% BSA
Preservative: 0.09% sodium azide
Storage Conditions: 4° C
Product Specific Information
By Western blot, MA5-11208 detects Human IgA (alpha) heavy chain at ~55kD in human, but not mouse samples.

By ELISA and WB, MA5-11208 does not react with other human Ig isotypes.
General Information
Thermo Scientific Pierce Anti-Human secondary antibodies are affinity-purified antibodies with well-characterized specificity for human immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
Product Images
  • Human IgA (Alpha heavy chain) Secondary Antibody (MA5-11208) in WB
    MA5-11208_WB_20131010122923.jpg

    Western Blot with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208) (MA5-11208_WB_20131010122923.jpg)

    Western Blot with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208)

    Western blot analysis of Human IgA (Alpha heavy chain) was performed by loading 25 ug of Daudi cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a Mouse anti-Human IgA (alpha-Heavy Chain) Secondary antibody (Product # MA5-11208) at a dilution of 1:100 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~60kDa.
  • Human IgA (Alpha heavy chain) Secondary Antibody (MA5-11208) in WB
    anti-IgA_Antibody_MA5-11208_Western-blot_1_20140711134927.jpg

    Western Blot with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208) (anti-IgA_Antibody_MA5-11208_Western-blot_1_20140711134927.jpg)

    Western Blot with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208)

    Western blot analysis of IgA was performed by loading the indicated amounts of purified Human IgA and 10ul of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with 5% BSA in TBST for at least 1 hour at room temperature. IgA heavy chain was detected at ~55kD using a mouse anti-human IgA monoclonal antibody (Product # MA5-11208) diluted to a concentration of 0.5ug/ml in blocking buffer, overnight at 4C overnight on a rocking platform. The blot was washed in TBST, and probed with an HRP-conjugated goat anti-mouse light chain secondary antibody at a dilution of 1:40,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico.
  • Human IgA (Alpha heavy chain) Secondary Antibody (MA5-11208) in WB
    anti-IgA_Antibody_MA5-11208_Western-blot_2_20140711134950.jpg

    Western Blot with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208) (anti-IgA_Antibody_MA5-11208_Western-blot_2_20140711134950.jpg)

    Western Blot with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208)

    Western blot analysis of IgA was performed by loading 0.5ug of purified Human IgA, Human IgG, Human IgM, and Mouse IgA, and 10ul of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membranes using the G2 Fast Blotter (Product # 62288), and blocked with 5% BSA in TBST for at least 1 hour at room temperature. IgA heavy chain was detected at ~55kD using a mouse anti-human IgA monoclonal antibody (Product # MA5-11208) diluted to a concentration of 0.5ug/ml in blocking buffer, overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse light chain secondary antibody diluted 1:40,000 for at least 30 minutes at room temperature (panel A). To verify the presence of all isotypes, a second blot was probed with a goat anti-human IgM + IgG +IgA polyclonal antibody (Product # 31128) at a concentration of 0.5ug/ml in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated mouse anti-goat IgG secondary antibody (Product # 31400) diluted 1:40,000 in blocking buffer for at least 30 minutes at room temperature (panel B). Chemiluminescent detection was performed using SuperSignal West Pico.
  • Human IgA (Alpha heavy chain) Secondary Antibody (MA5-11208) in WB
    anti-IgA_Antibody_MA5-11208_Western-blot_3_20140711135024.jpg

    Western Blot with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208) (anti-IgA_Antibody_MA5-11208_Western-blot_3_20140711135024.jpg)

    Western Blot with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208)

    Western blot analysis of IgA was performed by loading 25ug of various human tissue lysates, 0.5ug of Human IgA, 0.25ul of human serum, and 10ul of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with 5% BSA in TBST for at least 1 hour at room temperature. IgA heavy chain was detected at ~55kD using a mouse anti-human IgA monoclonal antibody (Product # MA5-11208) diluted to a concentration of 0.5ug/ml in blocking buffer, overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse light chain secondary antibody diluted 1:40,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico. *The band at ~25kD is a result of spill over from an adjacent lane.
  • Human IgA (Alpha heavy chain) Secondary Antibody (MA5-11208) in IHC
    MA5-11208_Immunohistochemistry.jpg

    Immunohistochemistry with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208) (MA5-11208_Immunohistochemistry.jpg)

    Immunohistochemistry with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208)

    Formalin-fixed, paraffin-embedded human tonsil stained with IgA antibody using peroxidase-conjugate and DAB chromogen. Note cytoplasmic staining of IgA secreting cells.
  • Human IgA (Alpha heavy chain) Secondary Antibody (MA5-11208) in FACS
    MA5-11208_FACS_Daudi_20130731120815.jpg

    Flow Cytometry with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208) (MA5-11208_FACS_Daudi_20130731120815.jpg)

    Flow Cytometry with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208)

    Flow cytometry analysis of Human IgA (alpha-Heavy Chain) in Daudi cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Mouse anti-Human IgA (alpha-Heavy Chain) Secondary antibody (Product # MA5-11208) at a dilution of 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
  • Human IgA (Alpha heavy chain) Secondary Antibody (MA5-11208) in FACS
    MA5-11208_FACS_Ramos_20130731120835.jpg

    Flow Cytometry with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208) (MA5-11208_FACS_Ramos_20130731120835.jpg)

    Flow Cytometry with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208)

    Flow cytometry analysis of Human IgA (alpha-Heavy Chain) in Ramos cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Mouse anti-Human IgA (alpha-Heavy Chain) Secondary antibody (Product # MA5-11208) at a dilution of 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
  • Human IgA (Alpha heavy chain) Secondary Antibody (MA5-11208) in ELISA
    anti-IgA_Antibody_MA5-11208_ELISA_1_20140711134846.jpg

    ELISA with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208) (anti-IgA_Antibody_MA5-11208_ELISA_1_20140711134846.jpg)

    ELISA with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208)

    Direct ELISA analysis of IgA was performed by coating wells of a 96-well plate with 100ul per well of Human IgA diluted in carbonate/bicarbonate buffer (Product # 28382), starting at a concentration of 5ug/ml and serially diluting 2-fold to a concentration of 5ng/ml, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer (Product # 37538), and incubated with 100ul per well of a mouse anti-human IgA monoclonal antibody (Product # MA5-11208) at a concentration of 1ug/ml for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondary antibody (Product # 31439) at a dilution of 1:8000 for 1 hour at room temperature. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.
  • Human IgA (Alpha heavy chain) Secondary Antibody (MA5-11208) in ELISA
    anti-IgA_Antibody_MA5-11208_ELISA_2_20140711134907.jpg

    ELISA with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208) (anti-IgA_Antibody_MA5-11208_ELISA_2_20140711134907.jpg)

    ELISA with anti-Human IgA (Alpha heavy chain) Monoclonal Secondary Antibody [GA01] (MA5-11208)

    Direct ELISA analysis of various immunoglobulin (Ig) isotypes was performed by coating wells of a 96-well plate with 100ul per well of Human IgA, Human IgG, Human IgM, or Mouse IgA diluted in carbonate/bicarbonate buffer (Product # 28382), starting at a concentration of 300ng/ml and serially diluting 2-fold to a concentration of 0.29ng/ml, overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer (Product # 37538), and incubated with 100ul per well of a mouse anti-human IgA monoclonal antibody (Product # MA5-11208) at a concentration of 1ug/ml for 1 hour at room temperature. The plate was washed, then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG Fc secondary antibody (Product # 31439) at a dilution of 1:8000 for 1 hour at room temperature. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 5 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid, and absorbances were read on a spectrophotometer at 450-550nm.
(This product is for In Vitro experimental use only. Not for resale without express authorization.)
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