Anti-Hsp40 antibody [EPR25331-69]

16 product images

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  Western blot - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  This antibody does not cross-react with human DNAJB5 (Hsp40 member 5) and DNAJB4 (Hsp40 member 4).

  All lanes: Western blot - Anti-Hsp40 antibody [EPR25331-69] (ab315992) at 1/1000 dilution

  Lane 1: His-tagged human DNAJB1 (Hsp40) recombinant protein at 10 ng

  Lane 2: His-tagged human DNAJB5 (Hsp40 member 5) recombinant protein at 20 ng

  Lane 3: His-tagged human DNAJB4 (Hsp40 member 4) recombinant protein at 40 ng

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Developed using the ECL technique.

  Observed band size: 38 kDa

  Exposure time: 180s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Hsp40 with ab315992 at 1/500 (0.992 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human colon. The section was incubated with ab315992 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labeling Hsp40 with ab315992 at 1/500 (0.992 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on human cervical cancer. The section was incubated with ab315992 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Hsp40 with ab315992 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on mouse colon. The section was incubated with ab315992 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Hsp40 with ab315992 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on rat colon. The section was incubated with ab315992 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Immunohistochemical analysis of paraffin-embedded (A) Wild-type HEK-293T (human epithelial cell line from embryonic kidney) cell pellet (B) DNAJB1 (Hsp40) knockout HEK-293T cell pellet tissue labeling Hsp40 with ab315992 at 1/5000 (0.099 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Positive staining on (A) Wild-type HEK-293T (human epithelial cell line from embryonic kidney) cell pellet, no staining on (B) DNAJB1 (Hsp40) knockout HEK-293T cell pellet. The section was incubated with ab315992 for 30 mins at room temperature.
  
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  Counterstained with Hematoxylin.

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

  Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• 

  Immunoprecipitation - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Hsp40 was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate with ab315992 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315992 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

  Lane 1: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
  
  Lane 2: ab315992 IP in HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
  
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab315992 in HEK-293 whole cell lysate

  All lanes: Immunoprecipitation - Anti-Hsp40 antibody [EPR25331-69] (ab315992) at 1/30 dilution

  All lanes: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 32s

• 

  Western blot - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  In Western blot, ab315992 was shown to bind specifically to DNAJB1 (Hsp40). Target of interest was observed at 38 kDa in wild-type HEK-293 cell lysates (lane 1) with no signal observed at this size in DNAJB1 (Hsp40) knockout cell line (lane 2, knockout cell line Human DNAJB1 (Hsp40) knockout HEK-293T cell line ab266344/ knockout cell lysate Human DNAJB1 (Hsp40) knockout HEK-293T cell lysate ab258400).

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] ab129002) staining at 1/10000 dilution.

  All lanes: Western blot - Anti-Hsp40 antibody [EPR25331-69] (ab315992) at 1/1000 dilution

  Lane 1: Wild-type HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

  Lane 2: DNAJB1 (Hsp40) knockout HEK-293 whole cell lysate at 20 µg

  Lane 3: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 4: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

  Lane 5: C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

  Lane 6: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Performed under reducing conditions.

  Observed band size: 38 kDa, 124 kDa

  Exposure time: 70s

• 

  Western blot - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  The identity of the higher MW band at approximately 120 kDa (in lane 2) is unknown.

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  All lanes: Western blot - Anti-Hsp40 antibody [EPR25331-69] (ab315992) at 1/1000 dilution

  Lane 1: Mouse testis tissue lysate at 20 µg

  Lane 2: Mouse colon tissue lysate at 20 µg

  Lane 3: Mouse liver tissue lysate at 20 µg

  Lane 4: Rat testis tissue lysate at 20 µg

  Lane 5: Rat colon tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 38 kDa, 36 kDa

  Exposure time: 70s

• 

  Western blot - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  The lanes 1 and 4 were developed using a high sensitivity ECL substrate.

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  Exposure time: Lane 1: 59 seconds, lane 2: 70 seconds, lanes 3-4: 180 seconds.

  All lanes: Western blot - Anti-Hsp40 antibody [EPR25331-69] (ab315992) at 1/1000 dilution

  Lane 1: Human colon cancer tissue lysate at 20 µg

  Lane 2: Human liver tissue lysate at 20 µg

  Lane 3: Human testis tissue lysate at 20 µg

  Lane 4: Human colon tissue lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Observed band size: 38 kDa, 36 kDa

• 

  Flow Cytometry (Intracellular) - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C6 (rat glial tumor glial cell) cells labelling Hsp40 with ab315992 at 1/500 dilution (0.1 ug)/(Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

• 

  Flow Cytometry (Intracellular) - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Hsp40 with ab315992 at 1/500 dilution (0.1 ug)/(Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

• 

  Flow Cytometry (Intracellular) - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized DNAJB1 (Hsp40) KO HEK293T (DNAJB1 (Hsp40) knockout human embryonic kidney epithelial cell, Left) / Parental HEK293T (Right) cells labelling Hsp40 with ab315992 at 1/500 dilution (0.1 ug)/(Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized DNAJB1 (Hsp40) KO HEK293T ( DNAJB1 (Hsp40) knockout human embryonic kidney epithelial cell) (Human DNAJB1 (Hsp40) knockout HEK-293T cell line ab266344) cells labelling Hsp40 with ab315992 at 1/100 (4.96 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

  Confocal image showing nuclear and cytoplasmic staining in parental HEK293T cells and no staining in DNAJB1 (Hsp40) KO HEK293T cells. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C6 (rat glial tumor glial cell) cells labelling Hsp40 with ab315992 at 1/100 (4.96 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

  Confocal image showing nuclear and cytoplasmic staining in C6 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Hsp40 antibody [EPR25331-69] (ab315992)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Hsp40 with ab315992 at 1/100 (4.96 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

  Confocal image showing nuclear and cytoplasmic staining in NIH/3T3 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.