Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23913-57] to PHF8 - BSA and Azide free
  • Suitable for: IHC-P, IP, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free
    See all PHF8 primary antibodies

  • Description

    Rabbit monoclonal [EPR23913-57] to PHF8 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, IP, WBmore details
    Unsuitable for: ChIP,Flow Cyt (Intra) or ICC

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK-293T whole cell lysate. Human cerebellum tissue lysate. Mouse and rat testis tissue lysate. HeLa and NIH/3T3, PC-12 whole cell lysate. His-tagged human PHF8 recombinant protein. IHC-P: Human and mouse testis, Rat colon tissue. IP: HeLa treated with 10µM MG-132 for 6 h.

  • General notes

    ab280896 is the carrier-free version of ab280887. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab280896 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Western blot - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Western blot - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    All lanes : Anti-PHF8 antibody [EPR23913-57] (ab280887) at 1/1000 dilution

    Lane 1 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 2 : U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 117 kDa
    Observed band size: 135/140 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab280887, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The observed MW is consistent with what has been described in the literature (PMID:27183383).

    Low expression: U-2 OS (PMID:27183383 )

    Lysates were made freshly and used in WB test immediately to minimize protein degradation.

    Exposure time: 81 seconds

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)

    This data was developed using ab280887, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human testis tissue labelling PHF8 with ab280887 at 1/200 (1.066 µg/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human testis. The section was incubated with ab280887 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunoprecipitation - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Immunoprecipitation - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)

    This data was developed using ab280887, the same antibody clone in a different buffer formulation.

    PHF8 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) treated with 10µM MG-132 for 6 hours, whole cell lysate 10 ug with ab280887 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280887 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) treated with 10µM MG-132 for 6 hours, whole cell lysate 10 ug

    Lane 2: ab280887 IP in HeLa treated with 10µM MG-132 for 6 hours, whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab280887 in HeLa treated with 10µM MG-132 for 6 hours, whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes

     

  • Western blot - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Western blot - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Anti-PHF8 antibody [EPR23913-57] (ab280887) at 1/1000 dilution + Human cerebellum tissue lysate at 20 µg

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

    Predicted band size: 117 kDa
    Observed band size: 135/140 kDa why is the actual band size different from the predicted?



    This data was developed using ab280887, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The observed MW is consistent with what has been described in the literature (PMID: 25047223).

    Exposure time: 158 seconds

     

  • Western blot - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Western blot - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    All lanes : Anti-PHF8 antibody [EPR23913-57] (ab280887) at 1/1000 dilution

    Lane 1 : Mouse testis tissue lysate
    Lane 2 : Rat testis tissue lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
    Lane 5 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 117 kDa
    Observed band size: 135/140 kDa why is the actual band size different from the predicted?



    This data was developed using ab280887, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The observed MW is consistent with what has been described in the literature (PMID:25047223).

    Lysates (lane 3-5) were made freshly and used in WB test immediately to minimize protein degradation.

    Exposure time: Lane 1-2: 158 seconds; Lane 3-5: 125 seconds

     

  • Western blot - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Western blot - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    All lanes : Anti-PHF8 antibody [EPR23913-57] (ab280887) at 1/5000 dilution

    Lane 1 : His-tagged human PHF8 recombinant protein
    Lane 2 : His-tagged human PHF2 recombinant protein

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 117 kDa
    Observed band size: 135/140 kDa why is the actual band size different from the predicted?



    This data was developed using ab280887, the same antibody clone in a different buffer formulation. 

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    This antibody has weak cross-reactivity with human PHF2. 

    These two recombinant proteins were made in house and expressed from E.coli expression systems.

    Exposure time: 8 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)

    This data was developed using ab280887, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue labelling PHF8 with ab280887 at 1/500 (1.066 µg/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse testis. The section was incubated with ab280887 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)

    This data was developed using ab280887, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded rat colon tissue labelling PHF8 with ab280887 at 1/500 (1.066 µg/ml) dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat colon. The section was incubated with ab280887 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)
    Anti-PHF8 antibody [EPR23913-57] - BSA and Azide free (ab280896)

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