Product Info Summary
SKU: | M01024-1 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Mouse |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Hsc70 Antibody Picoband™(monoclonal, 3B6)
View all HSPA8/HSC71/Hsc70 Antibodies
SKU/Catalog Number
M01024-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Hsc70 Antibody Picoband™ (monoclonal, 3B6) catalog # M01024-1. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Hsc70 Antibody Picoband™(monoclonal, 3B6) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01024-1)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
3B6
Isotype
Mouse IgG2b
Immunogen
E.coli-derived human Hsc70 recombinant protein (Position: Q520-A614). Human Hsc70 shares 98.9% amino acid (aa) sequence identity with both mouse and rat Hsc70.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M01024-1 is reactive to HSPA8 in Human, Mouse, Rat
Applications
M01024-1 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
71 kDa
Calculated molecular weight
70.898kDa
Background of HSPA8/HSC71/Hsc70
HSPA8 (heat shock 70kDa protein 8) also known as HSC70, HSC71, HSP73, HSPA10, FORMERLY, LAP1 or LPS-ASSOCIATED PROTEIN 1, is a heat shock protein that in humans is encoded by the HSPA8 gene. The HSPA8 gene contains 9 exons and spans 5 kb. The deduced HSPA8 protein has 646 amino acids and a predicted molecular mass of 70,899 Da. And the HSPA8 gene is mapped on 11q24.1. HSPA8 plays an important role in cells by transiently associating with nascent polypeptides to facilitate correct folding. HSP73 also functions as an ATPase in the disassembly of clathrin-coated vesicles during transport of membrane components through the cell. Rapid decay involves AU-rich binding protein AUF1, which complexes with heat-shock proteins HSC70 and HSP70, translation initiation factor EIF4G, and poly (A)-binding protein. In the absence of Il3, Hsc70 formed a complex with Hsp40 and Hip, and this complex, in association with Eif4g and Pabp, formed a high-stability complex with Bim mRNA that protected it from ribonucleases.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
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Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x106, Human, Mouse
Validation Images & Assay Conditions
Click image to see more details
Figure 10. Flow Cytometry analysis of HEPA1-6 cells using anti-Hsc70 antibody (M01024-1).
Overlay histogram showing HEPA1-6 cells stained with M01024-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Hsc70 Antibody (M01024-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Click image to see more details
Figure 1. Western blot analysis of Hsc70 using anti-Hsc70 antibody (M01024-1).
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 Hela whole cell lysates;
Lane 3: human Jurkat whole cell lysates;
Lane 4: human A549 whole cell lysates;
Lane 5: human HL-60 whole cell lysates;
Lane 6: rat PC-12 whole cell lysates;
Lane 7: human RAW264.7 whole cell lysates;
Lane 8: mouse NIH/3T3 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 mouse anti-Hsc70 antigen affinity purified monoclonal antibody (Catalog # M01024-1) 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-mouse 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 # EK1001) with Tanon 5200 system. A specific band was detected for Hsc70 at approximately 71KD. The expected band size for Hsc70 is at 71KD.
Click image to see more details
Figure 2. IHC analysis of Hsc70 using anti-Hsc70 antibody (M01024-1).
Hsc70 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hsc70 Antibody (M01024-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of Hsc70 using anti-Hsc70 antibody (M01024-1).
Hsc70 was detected in paraffin-embedded section of human rectal cancer (lymph node) tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hsc70 Antibody (M01024-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of Hsc70 using anti-Hsc70 antibody (M01024-1).
Hsc70 was detected in paraffin-embedded section of human rectal cancer (lymph node) tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hsc70 Antibody (M01024-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of Hsc70 using anti-Hsc70 antibody (M01024-1).
Hsc70 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hsc70 Antibody (M01024-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of Hsc70 using anti-Hsc70 antibody (M01024-1).
Hsc70 was detected in paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Hsc70 Antibody (M01024-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
Click image to see more details
Figure 7. IF analysis of Hsc70 using anti-Hsc70 antibody (M01024-1).
Hsc70 was detected in immunocytochemical section of A549 cells. 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 2μg/mL mouse anti-Hsc70 Antibody (M01024-1) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Mouse IgG (BA1141) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 8. Flow Cytometry analysis of A549 cells using anti-Hsc70 antibody (M01024-1).
Overlay histogram showing A549 cells stained with M01024-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Hsc70 Antibody (M01024-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Click image to see more details
Figure 9. Flow Cytometry analysis of K562 cells using anti-Hsc70 antibody (M01024-1).
Overlay histogram showing K562 cells stained with M01024-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Hsc70 Antibody (M01024-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For HSPA8 (Source: Uniprot.org, NCBI)
Gene Name
HSPA8
Full Name
Heat shock cognate 71 kDa protein
Weight
70.898kDa
Superfamily
heat shock protein 70 family
Alternative Names
Heat shock 70 kDa protein 8; heat shock 70kD protein 8; heat shock 70kDa protein 8; heat shock cognate 71 kDa protein; heat shock cognate protein 54; heat shock cognate protein, 71-kDa; HSC54; HSC70; HSC70heat shock 70kd protein 10; HSC71; HSP71; HSP73; HSP73constitutive heat shock protein 70; HSPA10; HSPA8; LAP1; lipopolysaccharide-associated protein 1; LPS-associated protein 1; MGC131511; MGC29929; NIP71; N-myristoyltransferase inhibitor protein 71 HSPA8 HEL-33, HEL-S-72p, HSC54, HSC70, HSC71, HSP71, HSP73, HSPA10, LAP-1, LAP1, NIP71 heat shock protein family A (Hsp70) member 8 heat shock cognate 71 kDa protein|LPS-associated protein 1|N-myristoyltransferase inhibitor protein 71|constitutive heat shock protein 70|epididymis luminal protein 33|epididymis secretory sperm binding protein Li 72p|heat shock 70kDa protein 8|heat shock 70kd protein 10|heat shock cognate protein 54|lipopolysaccharide-associated protein 1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on HSPA8, check out the HSPA8 Infographic
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In this infographic, you will see the following information for HSPA8: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Hsc70 Antibody Picoband™(monoclonal, 3B6) (M01024-1)
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