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- 供应商:
广州威佳科技有限公司
- 规格:
100ul
产品概况
| 货号 | BA0928 |
|---|---|
| 产品名称 | Anti-HSP70/HSPA1A/HSPA1B Antibody |
| 基因名 | HSPA1A/HSPA1B |
| 抗体来源 | Rabbit |
| 克隆 | Polyclonal |
| 抗体亚型 | Rabbit IgG |
| 分子量 | 70KD |
| 免疫原 | A synthetic peptide corresponding to a sequence at the N-terminus of human Hsp70(13-31aa TTYSCVGVFQHGKVEIIAN), identical to the related rat and mouse sequence. |
| 内容 | 500 ug/ml antibody with PBS ,0.02% NaN3 , 1 mg BSA and 50% glycerol. |
| 纯化方式 | Immunogen affinity purified. |
| 浓度 | 500 ug/ml |
| 产品形态 | Liquid |
| 保存条件 | 12 months from date of receipt,-20℃ as supplied. 6 months 2 to 8℃ after reconstitution. Avoid repeated freezing and thawing. |
| 背景资料 | The 70 kilodalton heat shock proteins(Hsp70s) are a family of ubiquitously expressed heat shock proteins. The Hsp70s are an important part of the cell's machinery for protein folding, and help to protect cells from stress. All of the Hsp70 proteins have three major functional domains: An N-terminal ATPase domain binds ATP(Adenosine triphosphate) and hydrolyzes it to ADP(Adenosine diphosphate); A substrate binding domain contains a groove with an affinity for neutral, hydrophobic amino acid residues; A C-terminal domain rich in alpha helical structure acts as a 'lid' for the substrate binding domain. By binding tightly to partially-synthesized peptide sequences(incomplete proteins), Hsp70 prevents them from aggregating and being rendered nonfunctional. And it also can act to protect cells from thermal or oxidative stress. Finally, Hsp70 seems to be able to participate in disposal of damaged or defective proteins. Interaction with CHIP(Carboxyl-terminus of Hsp70 Interacting Protein)–an E3 ubiquitin ligase–allows Hsp70 to pass proteins to the cell's ubiquitination and proteolysis pathways. |
| 研究类别 | 1. Tavaria M, Gabriele T, Kola I, Anderson RL (April 1996). "A hitchhiker's guide to the human Hsp70 family". Cell Stress Chaperones 1 (1): 23–8. 2. Morano KA (October 2007). "New tricks for an old dog: the evolving world of Hsp70". Ann. N. Y. Acad. Sci. 1113: 1–14. 3. Luders, J.; Demand, J.; Hohfeld, J. (2000), Journal of Biological Chemistry 275 (7): 4613–461, http://www.jbc.org/cgi/content/full/275/7/4613, retrieved on 2009-04-07 |
| Uniprot ID | HSPA1A/HSPA1B: P0DMV8/P0DMV9 |
| 推荐配套的二抗和检测试剂 | Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P). *Blocking peptide 可以联系我们购买。 |
产品应用细节
为了提供优质的抗体,博士德对每一批抗体都用没有转染过的细胞系和体细胞组织检测,以保证博士德出品的抗体有足够的亲和性足以和对应蛋白天然的表达含量起反应。
| 应用 | 稀释度* |
|---|---|
| Western blot(WB): | 1:500-2000 |
| Immunohistochemistry in paraffin section (IHC): | 1:50-400 |
| Immunohistochemistry in frozen section (IHC-F): | 1:50-400 |
| Immunocytochemistry/Immunofluorescence (ICC/IF): | 1:50-400 |
| Flow cytometry (FCM): | 1-3 μg/1x106 cells |
| (Boiling the paraffin sections in 10mM citrate buffer,pH6.0,or PH8.0 EDTA repair liquid for 20 mins is required for the staining of formalin/paraffin sections.) Optimal working dilutions must be determined by end user. | |
*稀释度需要用户自己调试,此处数据仅供参考。
**博士德提供高敏感的二抗和检测试剂盒。详情见相关产品推荐。
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[list_product_images]Figure 1. Western blot analysis of Hsp70 using anti- Hsp70 antibody (BA0928).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HEK293 whole cell lysates,
Lane 2: human Caco-2 whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human THP-1 whole cell lysates,
Lane 5: human U2OS whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- Hsp70 antigen affinity purified polyclonal antibody (Catalog # BA0928) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Hsp70 at approximately 70KD. The expected band size for Hsp70 is at 70KD.|Figure 2. IHC analysis of Hsp70 using anti-Hsp70 antibody (BA0928).
Hsp70 was detected in paraffin-embedded section of rat brain tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Hsp70 Antibody (BA0928) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 3. IHC analysis of Hsp70 using anti-Hsp70 antibody (BA0928).
Hsp70 was detected in paraffin-embedded section of mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Hsp70 Antibody (BA0928) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 4. IHC analysis of Hsp70 using anti-Hsp70 antibody (BA0928).
Hsp70 was detected in immunocytochemical section of HELA cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1μg/ml rabbit anti-Hsp70 Antibody (BA0928) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 5. IHC analysis of Hsp70 using anti-Hsp70 antibody (BA0928).
Hsp70 was detected in frozen section of rat intestine tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Hsp70 Antibody (BA0928) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 6. Western blot analysis of Hsp70 using anti- Hsp70 antibody (BA0928).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: rat liver tissue lysates,
Lane 3: rat spleen tissue lysates,
Lane 4: rat PC-12 whole cell lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse spleen tissue lysates,
Lane 8: mouse RAW246.7 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- Hsp70 antigen affinity purified polyclonal antibody (Catalog # BA0928) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Hsp70 at approximately 70KD. The expected band size for Hsp70 is at 70KD.|Figure 7.ICC analysis using anti-Hsp70 antibody (BA0928) was detected in immersion fixed HELA cell line . Cells were stained using the Dylight488-conjugated Anti-rabbit IgG Secondary Antibody (green)(Catalog#BA1127) and counterstained with DAPI (blue). |Figure 8.Flow cytometry analysis of 293T cell(1x106) DyLight 488 conjugated goat anti-rabbit IgG(blue) was used as secondary antibody.Isotype control antibody (Green line) was rabbit IgG DyLight 488. Unlabelled sample (Red line).[/list_product_images]
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HEK293 whole cell lysates,
Lane 2: human Caco-2 whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human THP-1 whole cell lysates,
Lane 5: human U2OS whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- Hsp70 antigen affinity purified polyclonal antibody (Catalog # BA0928) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Hsp70 at approximately 70KD. The expected band size for Hsp70 is at 70KD.|Figure 2. IHC analysis of Hsp70 using anti-Hsp70 antibody (BA0928).
Hsp70 was detected in paraffin-embedded section of rat brain tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Hsp70 Antibody (BA0928) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 3. IHC analysis of Hsp70 using anti-Hsp70 antibody (BA0928).
Hsp70 was detected in paraffin-embedded section of mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Hsp70 Antibody (BA0928) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 4. IHC analysis of Hsp70 using anti-Hsp70 antibody (BA0928).
Hsp70 was detected in immunocytochemical section of HELA cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1μg/ml rabbit anti-Hsp70 Antibody (BA0928) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 5. IHC analysis of Hsp70 using anti-Hsp70 antibody (BA0928).
Hsp70 was detected in frozen section of rat intestine tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Hsp70 Antibody (BA0928) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.|Figure 6. Western blot analysis of Hsp70 using anti- Hsp70 antibody (BA0928).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: rat liver tissue lysates,
Lane 3: rat spleen tissue lysates,
Lane 4: rat PC-12 whole cell lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse spleen tissue lysates,
Lane 8: mouse RAW246.7 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- Hsp70 antigen affinity purified polyclonal antibody (Catalog # BA0928) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Hsp70 at approximately 70KD. The expected band size for Hsp70 is at 70KD.|Figure 7.ICC analysis using anti-Hsp70 antibody (BA0928) was detected in immersion fixed HELA cell line . Cells were stained using the Dylight488-conjugated Anti-rabbit IgG Secondary Antibody (green)(Catalog#BA1127) and counterstained with DAPI (blue). |Figure 8.Flow cytometry analysis of 293T cell(1x106) DyLight 488 conjugated goat anti-rabbit IgG(blue) was used as secondary antibody.Isotype control antibody (Green line) was rabbit IgG DyLight 488. Unlabelled sample (Red line).[/list_product_images]
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Anti-Hsp70/HSPA1A/HSPA1B Antibody(BA0928-100ul)
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