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HA抗体琼脂糖珠(anti-H
A affinity gel)限时大特价1200元HA标签抗体偶联agarose beads
anti-HA antibody affinity gel
HA标签融合蛋白
50% slurry
Immunoprecipitation (IP), 40 ul of gel suspension (20μl of packed beads) for ~1000μg of crude total protein solution. Optimal dilutions/concentrations should be determined by the end user.
小鼠
all species
琼脂糖微球
单克隆
-20度
液体
Mouse IgG1
This monoclonal antibody is produced by immunizing mouses with a synthetic peptide (KLH-coupled) containing YPYDVPDYA
1ml
HA抗体琼脂糖珠(anti-HA affinity gel)限时大特价1200元(1ml规格)
微信:13817407320
产品详细信息如下:
Specificity: Anti-HA Agarose beads(Affinity gel) is a mouse monoclonal antibody that is covalently attached to agarose beads by hydrazide linkage. The antibody binds HA at the N-terminal, C-terminal and internal locations of fusion proteins.
Tested applications: Immunoprecipitation (IP),
40 ml of gel suspension (~20μl of packed beads) for ~1000μg of crude total protein solution.
Optimal dilutions/concentrations should be determined by the end user.
Reactivity: The antibody detects and captures HA-Tagged fusion proteins overexpressed in mammalian cells or E.coli.
Immunogen: This monoclonal antibody is produced by immunizing mouses with a synthetic peptide (KLH-coupled) containing YPYDVPDYA.
Form: The productis supplied as a 50% suspension in 50% glycerol with 10 mM sodium phosphate and 150 mM sodium chloride, pH 7.4, containing 0.02% (w/v) sodium azide
Binding capacity: 30-50 nmoles of HA-tagged fusion protein per 1 ml of settled gel.
Isotype: Mouse IgG1
Recommended IP Lysis buffer for mammalian cells: 50 mM Tris HCl, pH 7.4, with 150 mM NaCl, 1 mM EDTA, and 1% TRITON X-100.
Storage instruction:
The gel is supplied in 50% glycerol, so the gel can be stored at –20 °C for maximum stability. The unopened product is stable for one year when stored as indicated.
Attention: Agarose Beads should be resuspended well before used in IP
For used and recycled gel, the beads should be cleaned and stored in 50% glycerol with TBS or PBS buffer containing 0.02% sodium azide to protect the product. Stored at –20 °C for maximum stability or 4 °C for short term(1-2 weeks). Do not freeze in the absence of glycerol.
Immunoprecipitation(IP) protocol
Reagent | Effect | Notes |
Chaotropic agents (e.g., urea, guanidine HCl) | Denatures the immobilized anti HA antibody | Do not use any reagent that contains these types of components since it will denature the anti HA antibody on the gel and destroy its ability to bind the HA fusion proteins. Low concentrations of urea (1 M or less) can be used. |
Reducing agents (such as DTT, DTE, 2-mercaptoethanol) | Reduces the disulfide bridges holding the anti HA antibody chains together | Do not use any reagent that contains these types of components since it will reduce the disulfide linkages in the anti HA antibody on the gel and destroy its ability to bind the HA fusion proteins. |
Sodium dodecyl sulfate (SDS) | Denatures the immobilized anti HA antibody | Do not use any reagent that contains this detergent in the lysis and washing buffers since it will denature the anti HA antibody on the gel and destroy its ability to bind the HA fusion proteins. It is included in the sample loading buffer for removal of protein for immunoprecipitation, but the gel cannot be reused. |
Deoxycholate | Interferes with anti HA antibody binding to HA proteins | Do not use any reagent that contains this detergent since it will inhibit the anti HA antibody from binding to HA fusion proteins. |
Reagent | Effect | Notes |
TWEEN 20, 5% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 5%, but do not exceed. |
TRITON X-100 5% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 5%, but do not exceed. |
IGEPAL CA-630, 0.1% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 0.1%, but do not exceed. |
CHAPS, 0.1% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 0.1%, but do not exceed. |
Digitonin, 0.2% or less | Reduces nonspecific protein binding to the gel | May be used up to recommended concentration of 0.2%, but do not exceed. |
Sodium chloride, 1.0 M or less | Reduces nonspecific protein binding to the gel by reducing ionic interactions | May be used up to recommended concentration of 1.0 M, but do not exceed. |
0.1 M glycine HCl, pH 3.5 | Elutes HA protein from the gel | Do not leave the beads in glycine HCl for longer than 20 minutes. Longer incubation times will begin to denature the anti HA antibody |
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