FCM (Flow Cytometry)
(Figure 6. Flow Cytometry analysis of MOLT-4 cells using anti-HSPA4 antibody (AAA19794).Overlay histogram showing MOLT-4 cells stained with AAA19794 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HSPA4 Antibody (AAA19794, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
Rabbit HSPA4 Polyclonal Antibody | anti-HSPA4 antibody
Anti-HSPA4 Antibody Picoband
IHC: 2-5ug/ml, Human, Rat
ICC/IF: 5ug/ml, Human
FC/FACS (Fixed): 1-3ug/1x106 cells, Human
FCM (Flow Cytometry)
(Figure 6. Flow Cytometry analysis of MOLT-4 cells using anti-HSPA4 antibody (AAA19794).Overlay histogram showing MOLT-4 cells stained with AAA19794 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HSPA4 Antibody (AAA19794, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
FCM (Flow Cytometry)
(Figure 6. Flow Cytometry analysis of MOLT-4 cells using anti-HSPA4 antibody (AAA19794).Overlay histogram showing MOLT-4 cells stained with AAA19794 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HSPA4 Antibody (AAA19794, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
FCM (Flow Cytometry)
(Figure 5. Flow Cytometry analysis of 293T cells using anti-HSPA4 antibody (AAA19794).Overlay histogram showing 293T cells stained with AAA19794 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HSPA4 Antibody (AAA19794, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
FCM (Flow Cytometry)
(Figure 5. Flow Cytometry analysis of 293T cells using anti-HSPA4 antibody (AAA19794).Overlay histogram showing 293T cells stained with AAA19794 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HSPA4 Antibody (AAA19794, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.)
IHC (Immunohistochemistry)
(Figure 4. IHC analysis of HSPA4 using anti-HSPA4 antibody (AAA19794).HSPA4 was detected in a paraffin-embedded section of mouse ovary tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-HSPA4 Antibody (AAA19794) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-HSPA4 Antibody (AAA19794) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: human K562 whole cell lysates,Lane 3: rat brain tissue lysates,Lane 4: rat C6 whole cell lysates,Lane 5: mouse brain tissue lysates,Lane 6: mouse NIH/3T3 whole cell lysates,Lane 7: mouse Raw264.7 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-HSPA4 antigen affinity purified polyclonal antibody (#AAA19794) at 0.5ug/mL overnight at 4 degree 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (16 kDa.)
IHC (Immunohistochemistry)
(Figure 4. IHC analysis of HSPA4 using anti-HSPA4 antibody (AAA19794).HSPA4 was detected in a paraffin-embedded section of mouse ovary tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-HSPA4 Antibody (AAA19794) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-HSPA4 Antibody (AAA19794) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: human K562 whole cell lysates,Lane 3: rat brain tissue lysates,Lane 4: rat C6 whole cell lysates,Lane 5: mouse brain tissue lysates,Lane 6: mouse NIH/3T3 whole cell lysates,Lane 7: mouse Raw264.7 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-HSPA4 antigen affinity purified polyclonal antibody (#AAA19794) at 0.5ug/mL overnight at 4 degree 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (16 kDa.)
WB (Western Blot)
(Figure 1. Western blot analysis of HSPA4 using anti-HSPA4 antibody (AAA19794).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 30 ug of sample under reducing conditions.Lane 1: human 293T whole cell lysates,Lane 2: human K562 whole cell lysates,Lane 3: rat brain tissue lysates,Lane 4: rat C6 whole cell lysates,Lane 6: mouse brain tissue lysates,Lane 7: mouse NIH/3T3 whole cell lysates,Lane 8: mouse Raw264.7 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-HSPA4 antigen affinity purified polyclonal antibody (#AAA19794) at 0.25ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. A specific band was detected for HSPA4 at approximately 110 kDa. The expected band size for HSPA4 is at 96,14 kDa.)
WB (Western Blot)
(Figure 1. Western blot analysis of HSPA4 using anti-HSPA4 antibody (AAA19794).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 30 ug of sample under reducing conditions.Lane 1: human 293T whole cell lysates,Lane 2: human K562 whole cell lysates,Lane 3: rat brain tissue lysates,Lane 4: rat C6 whole cell lysates,Lane 6: mouse brain tissue lysates,Lane 7: mouse NIH/3T3 whole cell lysates,Lane 8: mouse Raw264.7 whole cell lysates.After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-HSPA4 antigen affinity purified polyclonal antibody (#AAA19794) at 0.25ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. A specific band was detected for HSPA4 at approximately 110 kDa. The expected band size for HSPA4 is at 96,14 kDa.)
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Product Notes
The HSPA4 hspa4 (Catalog #AAA19794) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-HSPA4 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's HSPA4 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS). WB: 0.1-0.25ug/ml, Human, Mouse, Rat IHC: 2-5ug/ml, Human, Rat ICC/IF: 5ug/ml, Human FC/FACS (Fixed): 1-3ug/1x106 cells, Human. Researchers should empirically determine the suitability of the HSPA4 hspa4 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "HSPA4, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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