Overview:
Product Name | HSP70 Antibody | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Description | Mouse Anti-Human Cell Membrane HSP70 Monoclonal IgG1 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Species Reactivity | Human, Mouse, Rat, Bovine, Pig | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Applications | WB, ICC/IF, FCM, FACS | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Antibody Dilution | WB (1:1000), ICC/IF (1:100), FACS (1:250); optimal dilutions for assays should be determined by the user. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Host Species | Mouse | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Immunogen Species | Human | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Immunogen | Human native HSP70 protein | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Concentration | 1 mg/ml | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Conjugates |
Alkaline Phosphatase, APC, ATTO 390, ATTO 488, ATTO 565, ATTO 594, ATTO 633, ATTO 655, ATTO 680, ATTO 700, Biotin, FITC, HRP, PE/ATTO 594, PerCP, RPE, Streptavidin, Unconjugated
StreptavidinProperties:
Streptavidin Datasheet BiotinProperties:
Biotin Datasheet HRP (Horseradish peroxidase)Properties:
HRP Datasheet AP (Alkaline Phosphatase)Properties:
AP Datasheet
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Properties
Storage Buffer | PBS pH 7.4, 50% glycerol, 0.09% Sodium Azide |
Storage Temperature | -20ºC |
Shipping Temperature | Blue Ice or 4ºC |
Purification | Protein G Purified |
Clonality | Monoclonal |
Clone Number | 1H11 |
Isotype | IgG1 |
Specificity | Binds to cell surface HSP70 without permeabilization of cell membrane in tumor cell lines. Does not bind to intracellular HSP70 at low concentrations. Detects a ~70 kDa band in Western Blot. |
Cite This Product | StressMarq Biosciences Cat# SMC-249, RRID: AB_2699628 |
Certificate of Analysis | A 1:250 dilution of SMC-249 was sufficient for detection of cell surface HSP70 in HCT116 cells using Fluorescence-activated cell sorting, applied in flow cytometry, with FITC as the fluorescent probe. |
Biological Description
Alternative Names | HSP70 1 Antibody, HSP70 2 Antibody, HSP70.1 Antibody, HSP72 Antibody, HSPA1 Antibody, HSPA1A Antibody, HSPA1B Antibody |
Research Areas | Cancer, Heat Shock, Cell Signaling, Chaperone Proteins, Protein Trafficking, Tumor Biomarkers |
Cellular Localization | Cell membrane |
Accession Number | NP_005336.3 |
Gene ID | 3303 |
Swiss Prot | P0DMV8 |
Scientific Background | HSP70 is a highly conserved protein that is ubiquitously expressed. It can be found in chloroplasts, eukaryotic cytosol, endoplasmic reticulum, and mitochondria, but also embedded in the cell membrane and in the extracellular space (1,2,3,4). Even though HSP70 is one of the most studied heat shock proteins, the export mechanism and method of membrane insertion are not fully understood. The majority of the proteins in the heat shock family lack a consensus signal for secretion via the ER-Golgi pathway (5). Researchers have found that HSP70 may be released from cells via exosomes or secretory vesicles (6). Although HSP70 is ubiquitously expressed, there is not much information about its presence on cell surface. The finding that HSP70 is localized on the cell surface of cancer cells but not normal cells suggests a conformational change of HSP70 in the lower pH environment characteristic of cancer cells (7). The presence of cell membrane embedded HSP70 has been found to increase the stability of cancer cells, thereby protecting tumors from environmental stress (8, 9). Anti-HSP70 antibody, clone 1H11 (Catalog No. SMC-249) is unique from other commercially available antibodies in that it can bind to the extracellular region of the cell membrane embedded HSP70 protein, allowing researchers to differentiate between membrane bound and intracellular HSP70 across cancer cells types. |
References |
1. Gribaldo, S. et al. (1999) J. Bacteriol. 181, 434-443. 2. Sharma, D. & Masison, D.C. (2009) Protein Pept. Lett. 16, 571-581. 3. Sharma, D. et al. (2009) PLoS. ONE.4, e6644. 4. Kampinga, H.H. & Craig, E.A. (2010) Nat. Rev. Mol. Cell Biol. 11, 579-592. 5. Nickel, W. & Seedorf, M. (2008) Annu. Rev. Cell Dev. Biol. 24, 287-308. 6. Multhoff, G. & Hightower, L.E. (1996) Cell Stress. Chaperones. 1, 167-176. 7. De Maio, A. (2011) Cell Stress. Chaperones. 16, 235-249. 8. Horvath, I. & Vigh, L. (2010) Nature. 463, 436-438. 9. Horvath, I., Multhoff, G., Sonnleitner, A., & Vigh, L. (2008) Biochim. Biophys. Acta 1778, 1653-1664. |
Product Images
Immunocytochemistry/Immunofluorescence analysis on non-permeabilized HCT116 cells using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC) showing cell membrane staining. Tissue: HCT116 cells. Species: Human. Fixation: 4% Formaldehyde. Permeabilization: None. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:100. Counterstain: Wheat germ agglutinin Texas red membrane marker; DAPI (blue) nuclear stain. Localization: Cell surface, cell membrane. (A) DAPI nuclear stain. (B) Wheat germ agglutinin Texas red. (C) HSP70 Antibody. (D) Composite. Courtesy of: Lawrence Hightower, Charles Giardina, and Didem Ozcan from University of Connecticut.
Fluorescence-activated cell sorting (FACS) using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC). Tissue: Jurkat E6.1. Species: Human. Unfixed/non-permeablized cells were incubated with (A) Anti-HSP70 control antibody, or (B) Anti-HSP70 antibody, clone 1H11 (SMC-249D-FITC), for 40 min at 4°C. Fixed/permeabilized cells were incubated with (C) Anti-HSP70 control antibody, or (D) Anti-HSP70 antibody, clone 1H11 (SMC-249D-FITC) for 40 min at 4°C. Analysis shows that Anti-HSP70 (SMC-249) can detect low levels of cell surface HSP70, and can fainlty detect intracellular HSP70. Courtesy of: Dr. Elyse Ireland, Institute of Medicine, University of Chester.
Western Blot analysis of Human Heat shocked HeLa cell lysates showing detection of HSP70 protein using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11 (SMC-249). Lane 1: Molecular Weight ladder (MW). Lane 2: HeLa cell lysates. Load: 20 µg. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:1000.
Immunocytochemistry/Immunofluorescence analysis on permeabilized HCT116 cells using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC) showing faint cell membrane and intracellular staining. Tissue: HCT116 cells. Species: Human. Fixation: 4% Formaldehyde. Permeabilization: 0.2% PBST. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:100. Counterstain: Wheat germ agglutinin Texas red membrane marker; DAPI (blue) nuclear stain. Localization: Cell surface, faint intracellular. (A) DAPI nuclear stain. (B) Wheat germ agglutinin Texas red. (C) HSP70 Antibody. (D) Composite. Courtesy of: Lawrence Hightower, Charles Giardina, and Didem Ozcan from University of Connecticut.
Fluorescence-activated cell sorting analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC). Tissue: HCT116 and HK2 cells. Species: Human. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:100 for 90 min at 4°C. Counterstain: Propidium Iodide nuclear stain. (A) HK2 cells unstained. (B) HK2 cells and HSP70 Antibody. (C) HCT116 cells unstained. (D) HCT116 cells and HSP70 Antibody. Shows that HSP70 Antibody binds to the cell surface of tumor cells but not non-tumor cells. Courtesy of: Lawrence Hightower, Charles Giardina, and Didem Ozcan from University of Connecticut.
Fluorescence-activated cell sorting analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC). Tissue: Jurkat E6.1, U937, MCF7 and HT29 cells . Species: Human. Fixation: No fixation. Permeabilization: None. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 20 µg/ml for 40 min at 4°C. Counterstain: Propidium Iodide nuclear stain at 2.5 µg/ml for 5 min at 4°C. Shows that binding of HSP70 Antibody is not cell-line specific. Courtesy of: Dr. Elyse Ireland, Institute of Medicine, University of Chester.
Product Citations (1)
Other Citations
Extracellular Hsp70 Enhances Mesoangioblast Migration via an Autocrine Signaling Pathway.
Barreca, M.M. et al. (2016) J Cell Physiol. 232(7):1845-1861.
PubMed ID: 27925208 Reactivity Mouse Applications: Protein Inhibition
FITC (Fluorescein) | ||
Overview:
FITC-Fluorescent-conjugate | Optical Properties: λex = 494 nm λem = 520 nm εmax = 7.3×104 Φf = 0.92 τfl = 5.0 ns Brightness = 67.2 Laser = 488 nm Filter set = FITC |
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1.直接用固体磷酸钠配制成50mM的磷酸钠溶液,再调pH到7.4;(我们试着用这个做了下,发现挂不上柱)
2.配置磷酸钠盐缓冲液:按NaH2PO4:Na2HPO4以19:81的摩尔比配制成pH7.4的缓冲液?(附一张百度出来的配方
)
3.如果是磷酸钠盐缓冲液,可以直接将50mM的NaH2PO4的水溶液用NaOH调成pH7.4吗?
再者,2和3这两个方法配制的磷酸钠盐缓冲液有什么区别?最终效果是一样的吗?如果不一样,有什么理论的知识支撑呢?个人感觉是分析化学中酸碱理论中的缓冲液那里的知识。求帮忙解答这些疑问。
另外,我还想问一下,pH对于Ni柱对His-tagged的蛋白的分离纯化影响大吗?是怎么影响的?谢谢大家了!
由弱酸及其盐、弱碱及其盐组成的混合溶液,能在一定程度上抵消、减轻外加强酸或强碱对溶液酸碱度的影响,从而保持溶液的pH值相对稳定。这种溶液称为缓冲溶液。
是否可以理解为纯化水得PH范围为6.3-7.6?能否直接用pH计测量?谢谢!
pH(1)=pKa+lg[c(CH₃COONa)/c(CH₃COOH)]=pKa=4.74
通HCl后,溶液是c(CH₃COOH)=0.2mol/L、c(NaCl)=0.1mol/L的混合溶液,溶液pH按照弱酸溶液pH的求法求.
c(H⁺)=√[Ka*c(CH₃COOH)]=√(10^-4.74*0.2)=0.00191(mol/L)(采用了近似公式)
pH(2)=-lg{c(H⁺)}=2.72
两个pH求得,那么pH的变化量也就可得了.pH的变化量=|pH(2)-pH(1)|=|2.72-4.74|=2.02
1)PH缓冲溶液作用原理和pH值
当往某些溶液中加入一定量的酸和碱时,有阻碍溶液pH变化的作用,称为缓冲作用,这样的溶液叫做缓冲溶液.弱酸及其盐的混合溶液(如HAc与NaAc),弱碱及其盐的混合溶液(如NH3·H2O与NH4Cl)等都是缓冲溶液.
由弱酸HA及其盐NaA所组成的缓冲溶液对酸的缓冲作用,是由于溶液中存在足够量的碱A-的缘故.当向这种溶液中加入一定量的强酸时,H离子基本上被A-离子消耗:
所以溶液的pH值几乎不变;当加入一定量强碱时,溶液中存在的弱酸HA消耗OH-离子而阻碍pH的变化.
2)PH缓冲溶液的缓冲能力
在缓冲溶液中加入少量强酸或强碱,其溶液pH值变化不大,但若加入酸,碱的量多时,缓冲溶液就失去了它的缓冲作用.这说明它的缓冲能力是有一定限度的.
缓冲溶液的缓冲能力与组成缓冲溶液的组分浓度有关.0.1mol·L-1HAc和0.1mol·L-1NaAc组成的缓冲溶液,比0.01mol·L-1HAc和0.01mol·L-1NaAc的缓冲溶液缓冲能力大.关于这一点通过计算便可证实.但缓冲溶液组分的浓度不能太大,否则,不能忽视离子间的作用.
组成缓冲溶液的两组分的比值不为1∶1时,缓冲作用减小,缓冲能力降低,当c(盐)/c(酸)为1∶1时△pH最小,缓冲能力大.不论对于酸或碱都有较大的缓冲作用.缓冲溶液的pH值可用下式计算:
此时缓冲能力大.缓冲组分的比值离1∶1愈远,缓冲能力愈小,甚至不能起缓冲作用.对于任何缓冲体系,存在有效缓冲范围,这个范围大致在pKaφ(或pKbφ)两侧各一个pH单位之内.
弱酸及其盐(弱酸及其共轭碱)体系pH=pKaφ±1
弱碱及其盐(弱碱及其共轭酸)体系pOH=pKbφ±1
例如HAc的pKaφ为4.76,所以用HAc和NaAc适宜于配制pH为3.76~5.76的缓冲溶液,在这个范围内有较大的缓冲作用.配制pH=4.76的缓冲溶液时缓冲能力最大,此时(c(HAc)/c(NaAc)=1.
3)PH缓冲溶液的配制和应用
为了配制一定pH的缓冲溶液,首先选定一个弱酸,它的pKaφ尽可能接近所需配制的缓冲溶液的pH值,然后计算酸与碱的浓度比,根据此浓度比便可配制所需缓冲溶液.
以上主要以弱酸及其盐组成的缓冲溶液为例说明它的作用原理、pH计算和配制方法.对于弱碱及其盐组成的缓冲溶液可采用相同的方法.
PH缓冲溶液在物质分离和成分分析等方面应用广泛,如鉴定Mg2离子时,可用下面的反应:
白色磷酸铵镁沉淀溶于酸,故反应需在碱性溶液中进行,但碱性太强,可能生成白色Mg(OH)2沉淀,所以反应的pH值需控制在一定范围内,因此利用NH3·H2O和NH4Cl组成的缓冲溶液,保持溶液的pH值条件下,进行上述反应.
:)
我在做一细菌不同酸碱度生长状况时,发现这些奇怪现象:pH=3的培养基灭菌(TSB液体培养基)灭菌后pH上升到到9.2!而原来pH=9.0的降到8.7(基本没多少变化),请问各位大侠,这是什么原因?
一般做不同酸碱度生长实验时,该如何才能防止pH在湿热灭菌后基本不变化?
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