Hemoglobin alpha Antibody (D-4)recommended for detection of Hemoglobin α of human origin by WB, IP, IF and ELISA

Hemoglobin α Antibody (D-4): sc-514378

Hemoglobin α Antibody (D-4) is rated 4.2 out of 5 by 6.
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Datasheets

    • Hemoglobin α Antibody (D-4) is a mouse monoclonal IgG1 (kappa light chain) provided at 200 µg/ml
    • raised against amino acids 62-142 of Hemoglobin α of human origin
    • Hemoglobin alpha Antibody (D-4) is recommended for detection of Hemoglobin α of human origin by WB, IP, IF and ELISA
    • Hemoglobin alpha Antibody (D-4) is available conjugated to agarose for IP; HRP for WB, IHC(P) and ELISA; and to either phycoerythrin or FITC for IF, IHC(P) and FCM
    • also available conjugated to Alexa Fluor® 488, Alexa Fluor® 546, Alexa Fluor® 594 or Alexa Fluor® 647 for WB (RGB), IF, IHC(P) and FCM, and for use with RGB fluorescent imaging systems, such as iBright™ FL1000, FluorChem™, Typhoon, Azure and other comparable systems
    • also available conjugated to Alexa Fluor® 680 or Alexa Fluor® 790 for WB (NIR), IF and FCM; for use with Near-Infrared (NIR) detection systems, such as LI-COR®Odyssey®, iBright™ FL1000, FluorChem™, Typhoon, Azure and other comparable systems
    • See m-IgGκ BP-HRP (mouse IgGκ binding protein-HRP), our highly recommended recombinant alternative to conventional secondary anti-mouse IgG reagents.
    • Contact our Technical Service Department (or your local Distributor) for more information on how to receive a FREE 10 µg sample of Hemoglobin α (D-4): sc-514378.
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Would like to know if there is any data regarding how does this antibody works with mouse-origin HBA and what kind of dilution could be suggested when using it to detect low abundant HBA (non-erythroid origin cells)

Asked by: LuísMonteiro
Thank you for your question. We do not recommend Hemoglobin α Antibody (D-4): sc-514378 for the detection of Hemoglobin α in murine samples. However, there is a publication that successfully used this antibody with murine samples in western blotting (PMID 29500821). The recommended starting dilution for human samples is 1:100 with a dilution range of 1:100-1:1000. For further information please contact technical service.
Answered by: SCBT Heidelberg Support
Date published: 2019-01-06

I want to run a Western blot using this Hemoglobin alpha monoclonal antibody using K-562 whole cell lysate. How much should I load on my gel?

Asked by: TinTin
We recommend loading ~50µg of cell lysate on your gel. You can find more about the recommended conditions in our Western blotting protocol: https://www.scbt.com/scbt/resources/protocols/western-immuno-blotting
Answered by: Technical Support 4
Date published: 2017-02-08
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Rated 5 out of 5 by from Worked well We used this antibody for WB of K562 cells. Loaded 20 ug protein, used primary Ab at 1/200 dilution, secondary at 1/1000 (anti-mouse-AP). Observed chemiluminescence band with lots of background (Immun-Star substrate), and a very clean band, no background using chromogenic substrate (BCIP/NBT).
Date published: 2018-08-22
Rated 4 out of 5 by from worked two month ago, i applied for this antibody and used it to detect the protein in Red blood cell sample, in generally, it worked, but instability.
Date published: 2017-03-04
Rated 5 out of 5 by from Good results in WB This monoclonal Hemoglobin alpha Antibody (D-4) yields specific and reliable results in Western blotting of human K-562, HEL 92.1.7, and TF-1 whole cell lysates.
Date published: 2017-01-27
Rated 1 out of 5 by from IHC (P): majority of erythrocytes unstained Formol-fixed (3,7% in tap water, 24h) normal human spleen specimens; antigen retrievals tested: 1) autoclaving in citrate buffer pH 6.0; 2) protease XIV; no pretreatment With all retrieval techniques the majority of erythrocytes remains unstained.
Date published: 2017-01-18
Rated 5 out of 5 by from Satisfactory Western Blot data of Hemoglobin Satisfactory Western Blot data of Hemoglobin expression in human plasma. -SCBT QC
Date published: 2015-02-10
Rated 5 out of 5 by from Nice Western Blot data of Hemoglobin expression Nice Western Blot data of Hemoglobin expression in K-562, HEL 92. -SCBT QC
Date published: 2013-11-25
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