SKU PA1214
Size 100μg/vial
Reactivity Human, Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications IHC, ICC, WB

Overview

Product Name Anti-Hsp70/HSPA1A/HSPA1B Antibody
SKU/Catalog Number PA1214
Storage & Handling At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for Heat shock 70 kDa protein 1A/1B(HSPA1A/HSPA1B) detection. Tested with WB, IHC-P, IHC-F, ICC in Human;Mouse;Rat.
Cite This Product Anti-Hsp70/HSPA1A/HSPA1B Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1214)
Host Rabbit
Contents/Buffer Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Form Lyophilized
Immunogen 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.
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunocytochemistry , 0.5-1μg/ml, Human, Mouse, Rat
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Mouse, Rat, Human
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

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), IHC(F) and ICC.

Images And Assay Conditions

/p/a/pa1214 1_1.jpg

Figure 1. Western blot analysis of Hsp70 using anti- Hsp70 antibody (PA1214).
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 # PA1214) 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.

/antibody/pa1214 2 IHC anti hsc70 antibody.jpg

Figure 2. IHC analysis of Hsp70 using anti-Hsp70 antibody (PA1214).
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 (PA1214) 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.

/antibody/pa1214 3 IHC anti hsc70 antibody.jpg

Figure 3. IHC analysis of Hsp70 using anti-Hsp70 antibody (PA1214).
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 (PA1214) 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.

/antibody/pa1214 4 IF anti hsc70 antibody.jpg

Figure 4. IHC analysis of Hsp70 using anti-Hsp70 antibody (PA1214).
Hsp70 was detected in immunocytochemical section of HELA cell. 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 (PA1214) 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.

/antibody/pa1214 5 IHC anti hsc70 antibody.jpg

Figure 5. IHC analysis of Hsp70 using anti-Hsp70 antibody (PA1214).
Hsp70 was detected in frozen section of rat intestine tissue. 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 (PA1214) 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.

/p/a/pa1214 6.jpg

Figure 6. Western blot analysis of Hsp70 using anti- Hsp70 antibody (PA1214).
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 # PA1214) 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.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id P0DMV8/P0DMV9
Gene Name HSPA8
Protein Name heat shock protein family A (Hsp70) member 1A/1B
Alternative Names Heat shock 70 kDa protein 1A; Heat shock 70 kDa protein 1B

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Research Areas Cancer, Chaperones, Heat Shock Proteins, Protein Trafficking, Signal Transduction, Tumor Biomarkers

*You can search these to find other products in these research areas.
Background 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.

Other Recommended Resources

Here are featured tools and databases that you might find useful.

Order Product (PA1214)

Promotion:

Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
Option Price
30ug sample size $99
100ug $280
100ug+Free HRP Secondary BA1054 $280
100ug+Free Biotin Secondary BA1003 $280

USD $280

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Troubleshooting

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Publications

Effects of intracerebral hemorrhage and subsequent minimally invasive hematoma aspiration on expression of apoptosisrelated genes in rats.
Effects of minocycline on the expression of NGF and HSP70 and its neuroprotection role following intracerebral hemorrhage in rats
Therapeutic efficacy of trehalose eye drops for treatment of murine dry eye induced by an intelligently controlled environmental system
Effect of pre-moxibustion on apoptosis and proliferation of gastric mucosa cells
Down-modulation of heat shock protein 70 and up-modulation of Caspase-3 during schisandrin B-induced apoptosis in human hepatoma SMMC-7721 cells

Customer Q&As

Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
A: You can find the immunogen sequence under "Immunogen" and clonality in the product name.
Q: What is the expected band size? Why is it different than the observed band size?
A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands

3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.

4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?
A: Some common names include but are not limited to lap1 antibody, hspa8 antibody