Anti-PDX1 antibody [RM1269] (ab323218)

Rabbit Recombinant Multiclonal PDX1 antibody. Suitable for IHC-P, WB, ICC/IF, IP, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [RM1269] (AB323218), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	WB	ICC/IF	IP	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Not recommended	Tested
Mouse	Tested	Tested	Tested	Tested	Tested
Rat	Tested	Expected	Expected	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes -
Species Mouse	Dilution info 1/50	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -
Species Mouse	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Target data

See full target information Pdx1

Function

Activates insulin and somatostatin gene transcription. Key regulator of islet peptide hormone expression but also responsible for the development of the pancreas, most probably by determining maturation and differentiation of common pancreatic precursor cells in the developing gut. As part of a PDX1:PBX1b:MEIS2b complex in pancreatic acinar cells is involved in the transcriptional activation of the ELA1 enhancer; the complex binds to the enhancer B element and cooperates with the transcription factor 1 complex (PTF1) bound to the enhancer A element. Binds the DNA sequence 5'-CC[CT]TAATGGG-3'.

Targets

PDX1, Pdx1

Alternative names

Ipf1, Pdx1, Pancreas/duodenum homeobox protein 1, Insulin promoter factor 1, Islet/duodenum homeobox 1, Somatostatin-transactivating factor 1, IPF-1, IDX-1, STF-1

Rabbit Recombinant Multiclonal PDX1 antibody. Suitable for IHC-P, WB, ICC/IF, IP, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal
Immunogens: The exact immunogen used to generate this antibody is proprietary information.
Clone number: RM1269
Purification technique: Affinity purification Protein A
Specificity: WB is not suitable for tissue lysates.
Not suitable for human IP and rat WB.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

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

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

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [RM1269] (ab323218)
Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling PDX1 with ab323218 at 1/2000 (0.263 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on mouse pancreas. The section was incubated with ab323218 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
Flow Cytometry (Intracellular) - Anti-PDX1 antibody [RM1269] (ab323218)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2(human hepatocellular carcinoma epithelial cell, Left) / DLD-1(human colorectal adenocarcinoma epithelial cell, Right) cells labelling PDX1 with ab323218 at 1/500 dilution (0.1ug) / Right 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.
Negative control:HepG2.
Flow Cytometry (Intracellular) - Anti-PDX1 antibody [RM1269] (ab323218)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3(mouse embryonic fibroblast, Left) / Beta-TC-6(mouse pancreas insulinoma beta cell, Right) cells labelling PDX1 with ab323218 at 1/500 dilution (0.1ug) / Right 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.
Negative control:NIH/3T3.
Immunoprecipitation - Anti-PDX1 antibody [RM1269] (ab323218)
PDX1 was immunoprecipitated from 0.35 mg Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate with ab323218 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323218 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: Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate

Lane 2: ab323218 IP in Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab323218 in Beta-TC-6 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-PDX1 antibody [RM1269] (ab323218) at 1/30 dilution
All lanes: Beta-TC-6 (mouse pancreas insulinoma beta 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: 8s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDX1 antibody [RM1269] (ab323218)
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling PDX1 with ab323218 at 1/100 (5.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: no staining on human kidney. The section was incubated with ab323218 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-PDX1 antibody [RM1269] (ab323218)
Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling PDX1 with ab323218 at 1/2000 (0.263 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on rat pancreas. The section was incubated with ab323218 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-PDX1 antibody [RM1269] (ab323218)
Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling PDX1 with ab323218 at 1/100 (5.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human pancreas. The section was incubated with ab323218 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
Immunocytochemistry/ Immunofluorescence - Anti-PDX1 antibody [RM1269] (ab323218)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized DLD-1(human colorectal adenocarcinoma epithelial cell) cells labelling PDX1 with ab323218 at 1/50 (10.5 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 2ug/ml dilution (Green).
Confocal image showing mainly nuclear staining in DLD-1 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Negative control: HepG2.

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 (Magenta). 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 2ug/ml dilution.
Immunocytochemistry/ Immunofluorescence - Anti-PDX1 antibody [RM1269] (ab323218)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Beta-TC-6 (mouse pancreas insulinoma beta cell) cells labelling PDX1 with ab323218 at 1/1000 (0.525 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 2ug/ml dilution (Green).
Confocal image showing nuclear staining in Beta-TC-6 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Negative control: NIH/3T3 (PMID: 25271154).

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 (Magenta). 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 2ug/ml dilution.
Western blot - Anti-PDX1 antibody [RM1269] (ab323218)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: NIH/3T3 (PMID: 25271154)
The molecular weight observed is consistent with what has been described in the literature (PMID: 12488243, PMID: 12947327, PMID: 16732298)
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-PDX1 antibody [RM1269] (ab323218) at 1/1000 dilution
Lane 1: Beta-TC-6 (mouse pancreas insinoma beta cell) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embryonic fibroblast) 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 1/20000 dilution
Observed band size: 46 kDa, 36 kDa
Exposure time: 37s
Western blot - Anti-PDX1 antibody [RM1269] (ab323218)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: HepG2, WI-38
The molecular weight observed is consistent with what has been described in the literature (PMID: 12488243, PMID: 12947327, PMID: 16732298)
This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.
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-PDX1 antibody [RM1269] (ab323218) at 1/1000 dilution
Lane 1: DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 40 µg
Lane 2: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 40 µg
Lane 3: WI-38 (human fetal lung fibroblast) whole cell lysate at 40 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Developed using the ECL technique.
Observed band size: 46 kDa, 36 kDa
Exposure time: 15s